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Volume 23,Issue 2,2016 Table of Contents
2016, 23(2):149-160.
DOI: 10.3872/j.issn.1007-385X.2016.02.001
Abstract:
Immunocyte therapy for tumor has drawn a great attention in recent years due to its significant effect. Immunocytes, including T cell, NK cell and DCs, play a key role in immune responses of anti-tumors and immunotherapy of tumors. Among them, the techique of chimeric antigen receptor (CAR) modified-T cell (CAR-T) and inhibitor therapy which reverses CTLA-4 and PD-1/PD-L1 and so on immune checkpoints of tumor immune suppressive function have respectively achieved exciting results in therapies of blood tumors, melanoma and other solid tumors. How to further improve the efficacy, to increase adaptive tumor diseases and to control immune related adverse reactions of the therapy could become the focus of future research. NK cell will also take advantages of CAR technique and inhibitors of immune checkpoints to further strengthen its role in the tumor therapy. How to enhance the curative effect of DCs as the first therapeutic tumor vaccine approved by FDA based on its confirmed safe and non-toxic side effects could become a hot point. In this paper, problems that need to be solved in the field were further analyzed and prospected with combination of recent advances in the immunocyte-therapy for tumor.
Immunocyte therapy for tumor has drawn a great attention in recent years due to its significant effect. Immunocytes, including T cell, NK cell and DCs, play a key role in immune responses of anti-tumors and immunotherapy of tumors. Among them, the techique of chimeric antigen receptor (CAR) modified-T cell (CAR-T) and inhibitor therapy which reverses CTLA-4 and PD-1/PD-L1 and so on immune checkpoints of tumor immune suppressive function have respectively achieved exciting results in therapies of blood tumors, melanoma and other solid tumors. How to further improve the efficacy, to increase adaptive tumor diseases and to control immune related adverse reactions of the therapy could become the focus of future research. NK cell will also take advantages of CAR technique and inhibitors of immune checkpoints to further strengthen its role in the tumor therapy. How to enhance the curative effect of DCs as the first therapeutic tumor vaccine approved by FDA based on its confirmed safe and non-toxic side effects could become a hot point. In this paper, problems that need to be solved in the field were further analyzed and prospected with combination of recent advances in the immunocyte-therapy for tumor.
2016, 23(2):161-167.
DOI: 10.3872/j.issn.1007-385X.2016.02.002
Abstract:
Objective:To construct a fusion protein prokaryotic expression vector of streptavidin (SA) and evolutionary conserved district (ECD) of Beclin1, to prepare PSMA targeting system the fusion protein coupling specific aptamer A10 of biotin-prostate specific membrane antigen (PSMA) and to identify its promotion effect on apoptosis and autophagy activities of PSMA+ prostatic cancer cells. Methods: Fusion SA-ECD gene was obtained by gene synthesis technology and then cloned into vector PUC57; after confirmation of correct synthesis, the SA-ECD gene was directly inserted into the prokaryotic expression vector PET30a; the fusion protein was expressed with inducement of IPTG, purified with Ni affinity gel chromatography column and identified with Western blotting. PSMA specific conjugate of PSMA, namely the targeting system, was prepared by mixing the biotin-A10 and the SA-ECD at the molar ratio of 4∶1; Combination of the biotin-A10 and the SA-ECD was confirmed by electrophoretic mobility shift assay (EMSA). Effects of the targeting system on apoptosis, expression of autophagy related proteins and viability of the prostatic cancer cells were detected by flow cytometry, Western blotting and trypan blue assays, respectively. Results:Double restriction enzyme digestion and gene sequencing assays verified that the recombinant plasmid PET30a-SA-ECD was successfully constructed; the SA-ECD fusion protein was efficiently expressed in E.coli with induction of IPTG and successfully purified by affinity gel chromatography. EMSA confirmed that SA-ECD could effectively bind biotin-A10. The targeting system could promote apoptosis and autophagy of the prostatic cancer cells, and inhibit its viability.Conclusion: The SA-ECD fusion protein was successfully expressed and purified, and the targeting system of biotin-A10 and SA-ECD could kill the PSMA+ prostatic cancer cells.
Objective:To construct a fusion protein prokaryotic expression vector of streptavidin (SA) and evolutionary conserved district (ECD) of Beclin1, to prepare PSMA targeting system the fusion protein coupling specific aptamer A10 of biotin-prostate specific membrane antigen (PSMA) and to identify its promotion effect on apoptosis and autophagy activities of PSMA+ prostatic cancer cells. Methods: Fusion SA-ECD gene was obtained by gene synthesis technology and then cloned into vector PUC57; after confirmation of correct synthesis, the SA-ECD gene was directly inserted into the prokaryotic expression vector PET30a; the fusion protein was expressed with inducement of IPTG, purified with Ni affinity gel chromatography column and identified with Western blotting. PSMA specific conjugate of PSMA, namely the targeting system, was prepared by mixing the biotin-A10 and the SA-ECD at the molar ratio of 4∶1; Combination of the biotin-A10 and the SA-ECD was confirmed by electrophoretic mobility shift assay (EMSA). Effects of the targeting system on apoptosis, expression of autophagy related proteins and viability of the prostatic cancer cells were detected by flow cytometry, Western blotting and trypan blue assays, respectively. Results:Double restriction enzyme digestion and gene sequencing assays verified that the recombinant plasmid PET30a-SA-ECD was successfully constructed; the SA-ECD fusion protein was efficiently expressed in E.coli with induction of IPTG and successfully purified by affinity gel chromatography. EMSA confirmed that SA-ECD could effectively bind biotin-A10. The targeting system could promote apoptosis and autophagy of the prostatic cancer cells, and inhibit its viability.Conclusion: The SA-ECD fusion protein was successfully expressed and purified, and the targeting system of biotin-A10 and SA-ECD could kill the PSMA+ prostatic cancer cells.
MOU Yakun
,
WANG Nan
,
GUO Dan
,
ZHAO Yangyang
,
LIU Xiaoyan
,
DONG Chengya
,
WANG Shuwei
,
WANG Lin
,
REN Qian
,
MA Xiaotong
2016, 23(2):168-174.
DOI: 10.3872/j.issn.1007-385X.2016.02.003
Abstract:
Objective:To investigate the expression of Twist-1 gene in patients with leukemia and tumor cell lines of hematopoietic system and to discuss the effect of its overexpression on proliferation and apoptosis of myeloid leukemic cells. Methods: Expressions of Twist-1 mRNA of hematopoietic tumor cell lines and PBMCs in patients with acute myeloid leukemia (AML), acute lymphoid leukemia (ALL) or chronic myeloid leukemia (CML) as well as in normal healthy human were detected by Real-time PCR. Lentiviral vector for overexperession of Twist-1 ( pCDH1-Twist-1) were constructed and transduced into myeloid leukemic K562, U937 and KG-1a cell lines. Effect of Twist-1 on proliferation, colony forming, cell cycle and apoptosis of the leukemic cells was evaluated by cell counting, colony forming, flow cytometry and Annexin V/PI assays. Results: Expressions of Twist-1 in patients with AML and CML were significantly higher than that in control group(all P<0.05), but there was no significant difference between all patients group and control group (P>005). The overexpression and interference analyses demonstrated that overexpression of Twist-1 in myeloid leukemic K562, U937 and KG-1a cell lines promoted proliferation and colony formation of the tumor cells, and inhibited apoptosis of the cells. Howevere, interfering expression of Twist-1 had the opposite effect. Conclusion: Twist-1 was overexpressed in myeloid leukemic K562, U937 and KG-1a cell lines and its overexpression could promote proliferation of the leukemia cells and inhibit their apoptosis.
Objective:To investigate the expression of Twist-1 gene in patients with leukemia and tumor cell lines of hematopoietic system and to discuss the effect of its overexpression on proliferation and apoptosis of myeloid leukemic cells. Methods: Expressions of Twist-1 mRNA of hematopoietic tumor cell lines and PBMCs in patients with acute myeloid leukemia (AML), acute lymphoid leukemia (ALL) or chronic myeloid leukemia (CML) as well as in normal healthy human were detected by Real-time PCR. Lentiviral vector for overexperession of Twist-1 ( pCDH1-Twist-1) were constructed and transduced into myeloid leukemic K562, U937 and KG-1a cell lines. Effect of Twist-1 on proliferation, colony forming, cell cycle and apoptosis of the leukemic cells was evaluated by cell counting, colony forming, flow cytometry and Annexin V/PI assays. Results: Expressions of Twist-1 in patients with AML and CML were significantly higher than that in control group(all P<0.05), but there was no significant difference between all patients group and control group (P>005). The overexpression and interference analyses demonstrated that overexpression of Twist-1 in myeloid leukemic K562, U937 and KG-1a cell lines promoted proliferation and colony formation of the tumor cells, and inhibited apoptosis of the cells. Howevere, interfering expression of Twist-1 had the opposite effect. Conclusion: Twist-1 was overexpressed in myeloid leukemic K562, U937 and KG-1a cell lines and its overexpression could promote proliferation of the leukemia cells and inhibit their apoptosis.
2016, 23(2):175-181.
DOI: 10.3872/j.issn.1007-385X.2016.02.004
Abstract:
Objective:To investigate function and maturation of DCs pulsed by α-GalCer and to explore influence of co-culture of CIK cells with α-GalCer-DCs on phenotype, proliferation activity and efficiency of killing hepatic carcinoma of the CIK cells. Methods: DCs and CIK cells were induced from monocytes that separated from human peripheral blood by density gradient centrifugation. The suspension and adherent monocytes were cultured ex vivo to generate DC and CIK cells respectively. The phenotypes of DCs pulsed by α-GalCer were detected by flow cytometry (FCM), and mRNA expression of related genes in DCs were detected by qRT-PCR. As co-culture of CIK cells with α-GalCer-DCs, surface markers on the DC and the CIK cells were detected by FCM. Proliferation multiple of the CIK cells was examined with trypan blue staining; expression levels of genes related with function of the CIK cells were measured by Real-time PCR; and effect of α-GalCer-DCs on killing HepG2 cells by the CIK cells was tested with CCK-8 kit. Results: CIK cells and mature DCs could be obtained from induction of multiple cytokines; Excitation of α-GalCer could promote DCs maturation, increase the positive expressions of CD80, CD86, CD83 and CD11c (P<0.05, P<0.01) and mRNA levels of surface chemokine receptors CCR-7 , IL-12, and IL-10 in the DCs (P<0.05). Co-cuture of the CIK cells with α-GalCer could significantly increase expression of CD3+CD56+ and proliferation activity of the CIK cells (P<0.05, P<0.01), and mRNA expression levels of INF-γ, IL-12, perforin and particle enzyme B (P<0.05, P<0.01); Killing effects of the CIK, the DC-CIK and the α-GalCer-DC-CIK cells on HepG2 cells were enhanced with increase of effector-target ratio. At the same effector-target ratio, α-GalCer-DC-CIK cells had the highest cytotoxicity effect on HepG2 cells (P<0.05).Conclusion:Pulse of α-GalCer could promote the maturation of DCs. Co-culture of the α-GalCer-DC with CIK cells could enhance proliferation and maturation of CIK cells, and enhance its activity to kill hepatic carcinoma cells. Therefore, the works might provide experimental and theoretical basis for application of the DC-CIK in immunotherapy of carcinoma.
Objective:To investigate function and maturation of DCs pulsed by α-GalCer and to explore influence of co-culture of CIK cells with α-GalCer-DCs on phenotype, proliferation activity and efficiency of killing hepatic carcinoma of the CIK cells. Methods: DCs and CIK cells were induced from monocytes that separated from human peripheral blood by density gradient centrifugation. The suspension and adherent monocytes were cultured ex vivo to generate DC and CIK cells respectively. The phenotypes of DCs pulsed by α-GalCer were detected by flow cytometry (FCM), and mRNA expression of related genes in DCs were detected by qRT-PCR. As co-culture of CIK cells with α-GalCer-DCs, surface markers on the DC and the CIK cells were detected by FCM. Proliferation multiple of the CIK cells was examined with trypan blue staining; expression levels of genes related with function of the CIK cells were measured by Real-time PCR; and effect of α-GalCer-DCs on killing HepG2 cells by the CIK cells was tested with CCK-8 kit. Results: CIK cells and mature DCs could be obtained from induction of multiple cytokines; Excitation of α-GalCer could promote DCs maturation, increase the positive expressions of CD80, CD86, CD83 and CD11c (P<0.05, P<0.01) and mRNA levels of surface chemokine receptors CCR-7 , IL-12, and IL-10 in the DCs (P<0.05). Co-cuture of the CIK cells with α-GalCer could significantly increase expression of CD3+CD56+ and proliferation activity of the CIK cells (P<0.05, P<0.01), and mRNA expression levels of INF-γ, IL-12, perforin and particle enzyme B (P<0.05, P<0.01); Killing effects of the CIK, the DC-CIK and the α-GalCer-DC-CIK cells on HepG2 cells were enhanced with increase of effector-target ratio. At the same effector-target ratio, α-GalCer-DC-CIK cells had the highest cytotoxicity effect on HepG2 cells (P<0.05).Conclusion:Pulse of α-GalCer could promote the maturation of DCs. Co-culture of the α-GalCer-DC with CIK cells could enhance proliferation and maturation of CIK cells, and enhance its activity to kill hepatic carcinoma cells. Therefore, the works might provide experimental and theoretical basis for application of the DC-CIK in immunotherapy of carcinoma.
2016, 23(2):182-187.
DOI: 10.3872/j.issn.1007-385X.2016.02.005
Abstract:
Objective:To evaluate the reversal effect of 4′-methylether-scutellarein (4′-M-S) on multidrug resistance of human choriocarcinoma in vivo, and investigate its mechanism. Methods: Drug resistant human choriocarcinoma cells were subcutaneously injected into left armpits of BALB/c nude mice to construct the subcutaneous exnograft model. When diameter of the xenografts reached 1.0 cm, the mice were randomly divided into four groups with ten mice in each, namely control grop, etoposide (VP16) group, 4′-M-S group and 4′-M-S+VP16 group. After the treatment for three weeks, growths of the exnograft tumors were dynamically observed and survival times of the mice recorded in the various groups, morphological changes of the tumor tissues were observed with optical and transmission electron microscopes and apoptosis rates of the exnograft cells detected with flow cytometry assay. Expression levels of MDR1 mRNA, LRP mRNA and their coding proteins, P-gp and LR, were detected and compared among the four groups with RT-PCR and Western blotting assays respectively. Results: After the treatments for three weeks, it was found that 4′-M-S has somewhat inhibitory effect on drug-resistant human choriocarcinoma in vivo, 4′-M-S+VP16 could apparently inhibit growth of the exnograft tumors in the nude mice with inhibition rate of 48.21%. Meanwhile, toxicity of the chemotherapy was reduced, life quality of mice with the exnograft tumors obviously improved and their mean survival times significantly prolonged. In the 4′-M-S+VP16 group, apoptosis rate of the xenograft cells was significantly increased, and expression levels of MDR1 mRNA, LRP mRNA and their coding proteins, P-gp and LR, in the exnograft tumors significantly decreased comparing with the other three groups (all P<0.05) . Conclusion: 4′-M-S could effectively reverse resistance of the human choriocarcinoma to the chemotherapy drug VP16, which might be related with 4′-M-S inducing apoptosis of the tumor cells and down-regulating expression of MDR1 mRNA, LRP mRNA and their coding proteins, P-gp and LR, in the exnograft tumors.
Objective:To evaluate the reversal effect of 4′-methylether-scutellarein (4′-M-S) on multidrug resistance of human choriocarcinoma in vivo, and investigate its mechanism. Methods: Drug resistant human choriocarcinoma cells were subcutaneously injected into left armpits of BALB/c nude mice to construct the subcutaneous exnograft model. When diameter of the xenografts reached 1.0 cm, the mice were randomly divided into four groups with ten mice in each, namely control grop, etoposide (VP16) group, 4′-M-S group and 4′-M-S+VP16 group. After the treatment for three weeks, growths of the exnograft tumors were dynamically observed and survival times of the mice recorded in the various groups, morphological changes of the tumor tissues were observed with optical and transmission electron microscopes and apoptosis rates of the exnograft cells detected with flow cytometry assay. Expression levels of MDR1 mRNA, LRP mRNA and their coding proteins, P-gp and LR, were detected and compared among the four groups with RT-PCR and Western blotting assays respectively. Results: After the treatments for three weeks, it was found that 4′-M-S has somewhat inhibitory effect on drug-resistant human choriocarcinoma in vivo, 4′-M-S+VP16 could apparently inhibit growth of the exnograft tumors in the nude mice with inhibition rate of 48.21%. Meanwhile, toxicity of the chemotherapy was reduced, life quality of mice with the exnograft tumors obviously improved and their mean survival times significantly prolonged. In the 4′-M-S+VP16 group, apoptosis rate of the xenograft cells was significantly increased, and expression levels of MDR1 mRNA, LRP mRNA and their coding proteins, P-gp and LR, in the exnograft tumors significantly decreased comparing with the other three groups (all P<0.05) . Conclusion: 4′-M-S could effectively reverse resistance of the human choriocarcinoma to the chemotherapy drug VP16, which might be related with 4′-M-S inducing apoptosis of the tumor cells and down-regulating expression of MDR1 mRNA, LRP mRNA and their coding proteins, P-gp and LR, in the exnograft tumors.
2016, 23(2):188-194.
DOI: 10.3872/j.issn.1007-385X.2016.02.006
Abstract:
Objective:To explore response of tumor-specific cytotoxic T lymphocyte (CTL) induced by dendritic cells (DCs) sensitized with exosomes that derived from heat-shocked gastric cancer cell. Methods: exosomes from mice murine foregastric cancer (MFC) cells (Exo), heat-shocked MFC cells (Exo/HS), and lysates (Lys) of MFC cells were prepared, respectively. Morphology of the exosomes was observed by electron microscopy and protein components of the exosomes were detected by Western blotting. DCs generated from murine bone marrow were cultured and sensitiaed with Exo/HS, Exo and Lys, and the prepared DCs vaccines were termed as DC-Exo/HS, DC-Exo, and DC-Lys, respectively. Also, DCs were sensitized with exosomes from heat-shocked MFC tumor cells that transfected with HSP70 siRNA. Phenotypes of DCs were detected by flow cytometry. Mice were immunized with DC-Exo/HS, DC-Exo, DC-Ly, DC, or PBS respectively; the proliferation of splenic T cells was measured by 3H-TdR and activity of spleen CTL was measured by LDH. MFC cells-bearing mouse models were constructed to examine immunotherapy effects of the DCs vaccines. Results: exosomes were confirmed as small membranous vesicles with electron microscopy. Detection of Western blotting showed that Exo/HS contained high level of HSP70. Flow cytometry demonstrated that the exosomes from heat-shocked MFC cells could significantly up-regulate expressions of MHC-Ⅱ, CD80, CD86 and CD40 molecules on the DCs, and interference with HSP70 siRNA could down-regulate expressions of MHC-Ⅱ, CD80, CD86 and CD40 molecules on the DCs sensitized with the exosomes from heat-shocked MFC tumor cells. Results of 3H-TdR showed that proliferation ability of T cells in DC-Exo/HS group was markedly enhanced as compared with those in DC-Exo, DC-Lys, DC and PBS groups (P<0.01). Results of LDH showed that higher level of CTL activity was induced in DC-Exo/HS group as compared with those in DC-Exo, DC-Lys, DC or PBS (P<0.01), and induced CTL activity in HSP70 siRNA group was significantly lower than that in control siRNA group (P<0.01). Results of the immunotherapies showed that inhibitive effect of DC-Exo/HS on tumor in the carcinoma-bearing mice was significantly better than those of all the other groups (P<001). Conclusion: DCs sensitized with exosomes that derived from heat-shocked gastric cancer cell could induce efficient antitumor immune response.
Objective:To explore response of tumor-specific cytotoxic T lymphocyte (CTL) induced by dendritic cells (DCs) sensitized with exosomes that derived from heat-shocked gastric cancer cell. Methods: exosomes from mice murine foregastric cancer (MFC) cells (Exo), heat-shocked MFC cells (Exo/HS), and lysates (Lys) of MFC cells were prepared, respectively. Morphology of the exosomes was observed by electron microscopy and protein components of the exosomes were detected by Western blotting. DCs generated from murine bone marrow were cultured and sensitiaed with Exo/HS, Exo and Lys, and the prepared DCs vaccines were termed as DC-Exo/HS, DC-Exo, and DC-Lys, respectively. Also, DCs were sensitized with exosomes from heat-shocked MFC tumor cells that transfected with HSP70 siRNA. Phenotypes of DCs were detected by flow cytometry. Mice were immunized with DC-Exo/HS, DC-Exo, DC-Ly, DC, or PBS respectively; the proliferation of splenic T cells was measured by 3H-TdR and activity of spleen CTL was measured by LDH. MFC cells-bearing mouse models were constructed to examine immunotherapy effects of the DCs vaccines. Results: exosomes were confirmed as small membranous vesicles with electron microscopy. Detection of Western blotting showed that Exo/HS contained high level of HSP70. Flow cytometry demonstrated that the exosomes from heat-shocked MFC cells could significantly up-regulate expressions of MHC-Ⅱ, CD80, CD86 and CD40 molecules on the DCs, and interference with HSP70 siRNA could down-regulate expressions of MHC-Ⅱ, CD80, CD86 and CD40 molecules on the DCs sensitized with the exosomes from heat-shocked MFC tumor cells. Results of 3H-TdR showed that proliferation ability of T cells in DC-Exo/HS group was markedly enhanced as compared with those in DC-Exo, DC-Lys, DC and PBS groups (P<0.01). Results of LDH showed that higher level of CTL activity was induced in DC-Exo/HS group as compared with those in DC-Exo, DC-Lys, DC or PBS (P<0.01), and induced CTL activity in HSP70 siRNA group was significantly lower than that in control siRNA group (P<0.01). Results of the immunotherapies showed that inhibitive effect of DC-Exo/HS on tumor in the carcinoma-bearing mice was significantly better than those of all the other groups (P<001). Conclusion: DCs sensitized with exosomes that derived from heat-shocked gastric cancer cell could induce efficient antitumor immune response.
2016, 23(2):195-200.
DOI: 10.3872/j.issn.1007-385X.2016.02.007
Abstract:
Objective:To explore whether arsenic trioxide (As2O3) negatively regulate function of tumor immune tolerance induced with inhibiting myeloid-derived suppressor cells (MDSCs) through inhibition of the MDSCs. Methods: Melanoma B16 cells and liver cancer H22 cells were subcutaneously injected into the C57BL/6 mice respectively to construct xenograft models. Growth of the xenografts was observed after treatment with As2O3. Immunophenotypes of MDSCs and other immune cells in spleen tissues of the mice with the xenograft, and effect of As2O3 (2 μmol/L) on the differentiation of MDSCs in the mouse models with B16 cells were detected by flow cytometry assay. the mouse models with B16 cells were randomly divided into two groups, namely As2O3 treatment group and control group. Changes of immunosuppressive activity of T cell mediated by MDSCs were detected with mixed lymphocytes reaction. Concentrations of TNF-α and IL-10 in serum of the mouse model with B16 cells and supernatant of MDSCs cultures were detected by ELISA. Results: As2O3 could inhibit growth of the xenografts in the mouse models with B16 and H22 cells, prolong survival periods of the mouse models with B16 cells, and significantly decrease percentages of MDSCs in spleen of the mouse models. After treatment with As2O3 (2 μmol/L ) in vitro for 5 d, the proportion of mature DCs (CD11c+/CD40+) in MDSCs of the mouse models with B16 cells was significantly higher than that of mice in control group (\[27.38±4.57\]% vs \[17.44±4.51\]%，P=0.0078). The immunosuppressive activity of MDSCs against T cells in the As2O3 treatment group was significantly lower than that in the control group (P=0.016), and the concentrations of TNF-α and IL-10 in mouse serum of As2O3 treatment group and supernatant of MDSCs cultures were significantly lower than that of the control group. Conclusion: As2O3 could induce maturation of MDSCs, down regulate its immunosuppressive activity and negatively regulate tumor suppression function of MDSCs.
Objective:To explore whether arsenic trioxide (As2O3) negatively regulate function of tumor immune tolerance induced with inhibiting myeloid-derived suppressor cells (MDSCs) through inhibition of the MDSCs. Methods: Melanoma B16 cells and liver cancer H22 cells were subcutaneously injected into the C57BL/6 mice respectively to construct xenograft models. Growth of the xenografts was observed after treatment with As2O3. Immunophenotypes of MDSCs and other immune cells in spleen tissues of the mice with the xenograft, and effect of As2O3 (2 μmol/L) on the differentiation of MDSCs in the mouse models with B16 cells were detected by flow cytometry assay. the mouse models with B16 cells were randomly divided into two groups, namely As2O3 treatment group and control group. Changes of immunosuppressive activity of T cell mediated by MDSCs were detected with mixed lymphocytes reaction. Concentrations of TNF-α and IL-10 in serum of the mouse model with B16 cells and supernatant of MDSCs cultures were detected by ELISA. Results: As2O3 could inhibit growth of the xenografts in the mouse models with B16 and H22 cells, prolong survival periods of the mouse models with B16 cells, and significantly decrease percentages of MDSCs in spleen of the mouse models. After treatment with As2O3 (2 μmol/L ) in vitro for 5 d, the proportion of mature DCs (CD11c+/CD40+) in MDSCs of the mouse models with B16 cells was significantly higher than that of mice in control group (\[27.38±4.57\]% vs \[17.44±4.51\]%，P=0.0078). The immunosuppressive activity of MDSCs against T cells in the As2O3 treatment group was significantly lower than that in the control group (P=0.016), and the concentrations of TNF-α and IL-10 in mouse serum of As2O3 treatment group and supernatant of MDSCs cultures were significantly lower than that of the control group. Conclusion: As2O3 could induce maturation of MDSCs, down regulate its immunosuppressive activity and negatively regulate tumor suppression function of MDSCs.
2016, 23(2):201-205.
DOI: 10.3872/j.issn.1007-385X.2016.02.008
Abstract:
Objective:To explore the expression characteristics of miR-129-5p (matured miR-129) in human glioma tissues, and to investigate whether the abnormal expression of miR-129-5p in human glioma tissues is related to methylation of its promoter. Methods: Twenty one tumor tissue specimens and twenty one non-tumor tissue specimens were respectively collected from patients with glioma and patients with brain trauma, who underwent surgery in Department of Neurosurgery, Xiangya Hospital during October to December, 2013. Expressions of miR-129-5 in human glioma tissues, nontumor brain tissues, glioma cell lines and normal glial cell lines were measured with Real-time PCR.Promoter methylation of miR-129-2 gene in glioma tissues was examined by methylation-specific PCR (MSP). Changes of promoter methylation level of miR-129-2 gene in the glioma tissues after treatment of methylation transferase inhibitor 5-aza-2′-deoxycitydine (5-Aza-dC) were analyzed with bisulfite-sequencing PCR (BSP). Results: The expressions of miR-129-5p in human giloma tissues and cells were significantly lower than those in normal brain tissues and cells (P<0.05). The promoter methylation rates of miR-129-2 gene in glioma tissues were significantly higher than those in normal brain tissues (P<0.05). After treatment of demethylation with 5-Aza-dC, the promoter methylation of miR-129-2 gene decreased and the expression of miR-129-5p significantly increased (P<0.05). Conclusion: miR-129-5p is significantly down-expressed in human giloma, promoter methylation of miR-129-2 is one of the mechanisms which result in low expression of miR-129-2 in human glioma.
Objective:To explore the expression characteristics of miR-129-5p (matured miR-129) in human glioma tissues, and to investigate whether the abnormal expression of miR-129-5p in human glioma tissues is related to methylation of its promoter. Methods: Twenty one tumor tissue specimens and twenty one non-tumor tissue specimens were respectively collected from patients with glioma and patients with brain trauma, who underwent surgery in Department of Neurosurgery, Xiangya Hospital during October to December, 2013. Expressions of miR-129-5 in human glioma tissues, nontumor brain tissues, glioma cell lines and normal glial cell lines were measured with Real-time PCR.Promoter methylation of miR-129-2 gene in glioma tissues was examined by methylation-specific PCR (MSP). Changes of promoter methylation level of miR-129-2 gene in the glioma tissues after treatment of methylation transferase inhibitor 5-aza-2′-deoxycitydine (5-Aza-dC) were analyzed with bisulfite-sequencing PCR (BSP). Results: The expressions of miR-129-5p in human giloma tissues and cells were significantly lower than those in normal brain tissues and cells (P<0.05). The promoter methylation rates of miR-129-2 gene in glioma tissues were significantly higher than those in normal brain tissues (P<0.05). After treatment of demethylation with 5-Aza-dC, the promoter methylation of miR-129-2 gene decreased and the expression of miR-129-5p significantly increased (P<0.05). Conclusion: miR-129-5p is significantly down-expressed in human giloma, promoter methylation of miR-129-2 is one of the mechanisms which result in low expression of miR-129-2 in human glioma.
9 Inhibitory effect of PAMAM-NK4 nanocomposite on the growth of breast cancer xenografts in nude mice
YAN Yan
,
LIU Minfeng
,
SUN Shiqiang
,
LIU Runqi
,
DONG Jianyu
,
GUO Zhaoze
,
GUO Jingyun
,
WANG Hengyu
,
YE Changsheng
,
LI Wenji
2016, 23(2):206-211.
DOI: 10.3872/j.issn.1007-385X.2016.02.009
Abstract:
Objective:To explore the inhibitory effect of polyamidoamine dendrimer (PAMAM)-mediated NK4 gene on the growth of breast cancer cell lines MDA-MB-231 and MCF-7, and its therapeutic effect on MDA-MB-231 cell transplantation tumor in nude mouse model. Methods: PAMAM-NK4 nanoparticles and blank plamids were prepared and respectively transfected into MDA-MB-231 and MCF-7 cells as the experiment group (PAMAM-NK4 group) and the control group (PAMAM group). MTT, Western blotting and flow cytometry assays were used to examine effect of the PAMAM-NK4 transfection on proliferation of the cells, expression of NK4 protein and apoptosis of the cells, respectively. Forty female nude mice were subcutaneous injected with human breast cancer MDA-MB-231 cell to establish nude mouse model with the xenograft tumor. The mice were randomly divided into 4 groups with 10 mice in each. Mice of the blank control group were subcutaneously injected with 0.2 ml of 0.9% NaCl surrounding the xenograft tumor, those of the blank plasmid group with 0.2 ml of plasmid solution containing 100 μg PAMAM-LacZ, those of the PAMAM-NK4 group with 0.2 ml of plasmid solution containing 100 μg PAMAM-NK4 and those of the positive control group with 0.2 ml of solution containing 100 μg adriamycin, respectively. All of the nude mice were continuously injected for 7 d and then sacrificed on 30th day. The xenograft tumors were removed completely to measure their weights and sizes. Expressions of the NK4 protein in the xenograft tissues were detected by Western blotting assay. Results: The transfection of PAMAM-NK4 nanocomposites, could result in stable expression of the NK4 protein, and inhibition of the cell proliferation and promotion of the cell apoptosis. The nude mouse models with human breast carcinoma xenogragts were successfully constructed. The volumes and weights of the carcinoma xenografts in the PAMAM-NK4 and the positive control groups were significantly smaller than those in the blank control group (P<0.05) with a fairly good safety. The expression of NK4 protein in the PAMAM-NK4 group significantly increased, comparing with that in the blank control group (P<0.05). Conclution: PAMAM-NK4 nanocomposites could inhibit growth of the breast cancer MDA-MB-231 and MCF-7 cells, and have therapeutic effect on the human breast carcinoma xenografts in nude mouse model.
Objective:To explore the inhibitory effect of polyamidoamine dendrimer (PAMAM)-mediated NK4 gene on the growth of breast cancer cell lines MDA-MB-231 and MCF-7, and its therapeutic effect on MDA-MB-231 cell transplantation tumor in nude mouse model. Methods: PAMAM-NK4 nanoparticles and blank plamids were prepared and respectively transfected into MDA-MB-231 and MCF-7 cells as the experiment group (PAMAM-NK4 group) and the control group (PAMAM group). MTT, Western blotting and flow cytometry assays were used to examine effect of the PAMAM-NK4 transfection on proliferation of the cells, expression of NK4 protein and apoptosis of the cells, respectively. Forty female nude mice were subcutaneous injected with human breast cancer MDA-MB-231 cell to establish nude mouse model with the xenograft tumor. The mice were randomly divided into 4 groups with 10 mice in each. Mice of the blank control group were subcutaneously injected with 0.2 ml of 0.9% NaCl surrounding the xenograft tumor, those of the blank plasmid group with 0.2 ml of plasmid solution containing 100 μg PAMAM-LacZ, those of the PAMAM-NK4 group with 0.2 ml of plasmid solution containing 100 μg PAMAM-NK4 and those of the positive control group with 0.2 ml of solution containing 100 μg adriamycin, respectively. All of the nude mice were continuously injected for 7 d and then sacrificed on 30th day. The xenograft tumors were removed completely to measure their weights and sizes. Expressions of the NK4 protein in the xenograft tissues were detected by Western blotting assay. Results: The transfection of PAMAM-NK4 nanocomposites, could result in stable expression of the NK4 protein, and inhibition of the cell proliferation and promotion of the cell apoptosis. The nude mouse models with human breast carcinoma xenogragts were successfully constructed. The volumes and weights of the carcinoma xenografts in the PAMAM-NK4 and the positive control groups were significantly smaller than those in the blank control group (P<0.05) with a fairly good safety. The expression of NK4 protein in the PAMAM-NK4 group significantly increased, comparing with that in the blank control group (P<0.05). Conclution: PAMAM-NK4 nanocomposites could inhibit growth of the breast cancer MDA-MB-231 and MCF-7 cells, and have therapeutic effect on the human breast carcinoma xenografts in nude mouse model.
2016, 23(2):212-217.
DOI: 10.3872/j.issn.1007-385X.2016.02.010
Abstract:
Objective:Gene expression profile is an important pattern to discover new uses of the existing drugs. Application value of the gene expression profile in screening of candidate drugs for colon cancer was discussed. Methods: First of all, data set of gene expression profile for colon cancer (No. GSE41258) was obtained from GEO data bank of PubMed. Fifty-three paired data of colon cancer and colonic mucosal tissues were selected from Memorial Sloan-Kettering Cancer Center of the United States, and their qualities analyzed with RMA express software. Then a gene expression profile for characteristics of colon cancer was established with Dchip software. Finally candidate compounds for therapy of colon cancer were screened by Connectivity Map (CMAP) and confirmed by experiments. Results: Thirty-six paired samples were selected through analysis of microarray chips for further research. A gene expression profile for characteristics of colon cancer was composed of 371 genes which include 94 up-regulation genes and 277 down-regulation genes. With analysis of CMAP, 10 candidate compounds, including vorinostat, tanespimycin (17-AAG) etc, were screened out, in which 17-AAG with high enrichment scores was selected for the validation. By the following experiments, it was confirmed that 17-AAG can effectively inhibit proliferation of colon cancer SW480 and HT29 cell lines. After treatment of 17-AAG for 24 and 48h, the IC50 values of SW480 cell line were 0.73 μmol/L and 0.41 μmol/L respectively, while the IC50 values of HT29 cell line was 0.72 μmol/L and 0.5 μmol/L respectively. Analysis of flow cytometry assay showed that 17-AAG could block cell cycle of the colon cancer cells at G1 phase. Conclusion: Multiple candidate therapeutic compounds for colon cancer were identified based on a pattern of gene expression profiles. A pattern of gene expression profiles might be of an important significance in screening of candidate drugs.
Objective:Gene expression profile is an important pattern to discover new uses of the existing drugs. Application value of the gene expression profile in screening of candidate drugs for colon cancer was discussed. Methods: First of all, data set of gene expression profile for colon cancer (No. GSE41258) was obtained from GEO data bank of PubMed. Fifty-three paired data of colon cancer and colonic mucosal tissues were selected from Memorial Sloan-Kettering Cancer Center of the United States, and their qualities analyzed with RMA express software. Then a gene expression profile for characteristics of colon cancer was established with Dchip software. Finally candidate compounds for therapy of colon cancer were screened by Connectivity Map (CMAP) and confirmed by experiments. Results: Thirty-six paired samples were selected through analysis of microarray chips for further research. A gene expression profile for characteristics of colon cancer was composed of 371 genes which include 94 up-regulation genes and 277 down-regulation genes. With analysis of CMAP, 10 candidate compounds, including vorinostat, tanespimycin (17-AAG) etc, were screened out, in which 17-AAG with high enrichment scores was selected for the validation. By the following experiments, it was confirmed that 17-AAG can effectively inhibit proliferation of colon cancer SW480 and HT29 cell lines. After treatment of 17-AAG for 24 and 48h, the IC50 values of SW480 cell line were 0.73 μmol/L and 0.41 μmol/L respectively, while the IC50 values of HT29 cell line was 0.72 μmol/L and 0.5 μmol/L respectively. Analysis of flow cytometry assay showed that 17-AAG could block cell cycle of the colon cancer cells at G1 phase. Conclusion: Multiple candidate therapeutic compounds for colon cancer were identified based on a pattern of gene expression profiles. A pattern of gene expression profiles might be of an important significance in screening of candidate drugs.
2016, 23(2):218-222.
DOI: 10.3872/j.issn.1007-385X.2016.02.011
Abstract:
Objective:To investigate the effect of siRNA targeted silencing CDX2 gene combined with vincristine (VCR) on the proliferation and apoptosis of leukemia K562 cells. Methods: Specific siRNA sequence targeted silencing CDX2 gene (CDX2-siRNA-921) and its negative control sequence (CDX2-siRNA-NC) were designed and transfected into K562 Cells respectively. At the same time, untransfected K562 cells were used as a control group. The effect of CDX2-siRNA-921 transfection on the expressions of CDX2，BCR-ABL mRNA and CDX2 protein was detected by Real-time PCR and Western blotting respectively. After action of siRNA and VCR alone or in combination, proliferation inhibition rate and apoptosis rate of the K562 cells were respectively detected by MTT and flow cytometry assays. Results: Compared with the control group, CDX2-siRNA-921 significantly decreased the expressions of targeting gene CDX2 and BCR-ABL mRNA, and CDX2 protein (all P<0.05). But the expressions of CDX2 and BCR-ABL mRNA, and CDX2 protein in CDX2-siRNA-NC group had nonsignificant changes. Compared with CDX2-siRNA-NC、VCR and VCR+CDX2-siRNA-NC groups, the proliferation inhibition rate of the K562 cells significantly decreased and apoptosis rate of the cells significantly increased in VCR+CDX2-siRNA-921 group (all P<0.05). Conclusions: CDX2-siRNA-921 could obviously decrease the expressions of mRNA and protein of CDX2 gene and down-regulate the expression of BCR-ABL fusion gene in the K562 cells, as well as enhance the effects of VCR on proliferation inhibition and inducing apoptosis of K562 cells.
Objective:To investigate the effect of siRNA targeted silencing CDX2 gene combined with vincristine (VCR) on the proliferation and apoptosis of leukemia K562 cells. Methods: Specific siRNA sequence targeted silencing CDX2 gene (CDX2-siRNA-921) and its negative control sequence (CDX2-siRNA-NC) were designed and transfected into K562 Cells respectively. At the same time, untransfected K562 cells were used as a control group. The effect of CDX2-siRNA-921 transfection on the expressions of CDX2，BCR-ABL mRNA and CDX2 protein was detected by Real-time PCR and Western blotting respectively. After action of siRNA and VCR alone or in combination, proliferation inhibition rate and apoptosis rate of the K562 cells were respectively detected by MTT and flow cytometry assays. Results: Compared with the control group, CDX2-siRNA-921 significantly decreased the expressions of targeting gene CDX2 and BCR-ABL mRNA, and CDX2 protein (all P<0.05). But the expressions of CDX2 and BCR-ABL mRNA, and CDX2 protein in CDX2-siRNA-NC group had nonsignificant changes. Compared with CDX2-siRNA-NC、VCR and VCR+CDX2-siRNA-NC groups, the proliferation inhibition rate of the K562 cells significantly decreased and apoptosis rate of the cells significantly increased in VCR+CDX2-siRNA-921 group (all P<0.05). Conclusions: CDX2-siRNA-921 could obviously decrease the expressions of mRNA and protein of CDX2 gene and down-regulate the expression of BCR-ABL fusion gene in the K562 cells, as well as enhance the effects of VCR on proliferation inhibition and inducing apoptosis of K562 cells.
TANG Haibin
,
MA Yueyun
,
SU Mingquan
,
LI Rui
,
CHANG Liang
,
GAO Meng
,
FU Xiaorui
,
YANG Yuqi
,
HAO Xiaoke
2016, 23(2):223-229.
DOI: 10.3872/j.issn.1007-385X.2016.02.012
Abstract:
Objective:To investigate roles of cAMP-response element binding protein (CREB) and phosphorglated CREB (p-CREB) in proliferation of prostate cancer cells.Methods:Tissue microarray technology was adopted to examine the expression levels of CREB and p-CREB in normal prostate tissue, hyperplasia prostate tissue and prostate cancer tissues at different grades. Immuno fluorescence staining and Western blotting were performed to detect the expression levels of CREB and p-CREB in human normal prostatic stromal immortalized WPMY-1 cell line, prostate cancer PC3 and LNCap cell lines. Recombinant human plasmid CREB was constructed and transfected into the PC3 and LNCap cells. Efficiency of the transfection and proliferation changes of the cells were verified by Western blotting and CCK-8 kit respectively. Results: Expression rates of CREB and p-CREB in prostate cancer tissue were significantly higher than those in normal prostate tissue (96% vs 50% and 88% vs 40%, P<0.05) and hyperplasia prostate tissue (96% vs 80% and 88% vs 60%, P<0.05). Expression levels of CREB and p-CREB proteins in the prostate cancer PC3 and LNCap cell lines were significantly higher than those in the normal prostatic stromal immortalized WPMY-1 cell line \[(5.10±0.62 vs 2.31±040), (7.5±0.83 vs 2.31±0.40), (4.31±0.54 vs 0.02±0.38), (6.15±0.69 vs 2.02±0.38), all P<0.05\]. Transfection with recombined CREB plasmid could up-regulate the expression of proliferating cell nuclear antigen (PCNA) in the prostate tumor PC3 and LNCap cells (all P<0.05), and proliferation levels of the PC3 and LNCap cells significantly increased after the transfection (P<0.05). Conclusion: The CREB and p-CREB were highly expressed in human prostate cancer tissues and, the prostate cancer PC3 and LNCap cells cultured in vitro. In addition, the CREB and p-CREB could up-regulate the expressions of proliferation-related gene PCNA, and raise proliferation level of the cells.
Objective:To investigate roles of cAMP-response element binding protein (CREB) and phosphorglated CREB (p-CREB) in proliferation of prostate cancer cells.Methods:Tissue microarray technology was adopted to examine the expression levels of CREB and p-CREB in normal prostate tissue, hyperplasia prostate tissue and prostate cancer tissues at different grades. Immuno fluorescence staining and Western blotting were performed to detect the expression levels of CREB and p-CREB in human normal prostatic stromal immortalized WPMY-1 cell line, prostate cancer PC3 and LNCap cell lines. Recombinant human plasmid CREB was constructed and transfected into the PC3 and LNCap cells. Efficiency of the transfection and proliferation changes of the cells were verified by Western blotting and CCK-8 kit respectively. Results: Expression rates of CREB and p-CREB in prostate cancer tissue were significantly higher than those in normal prostate tissue (96% vs 50% and 88% vs 40%, P<0.05) and hyperplasia prostate tissue (96% vs 80% and 88% vs 60%, P<0.05). Expression levels of CREB and p-CREB proteins in the prostate cancer PC3 and LNCap cell lines were significantly higher than those in the normal prostatic stromal immortalized WPMY-1 cell line \[(5.10±0.62 vs 2.31±040), (7.5±0.83 vs 2.31±0.40), (4.31±0.54 vs 0.02±0.38), (6.15±0.69 vs 2.02±0.38), all P<0.05\]. Transfection with recombined CREB plasmid could up-regulate the expression of proliferating cell nuclear antigen (PCNA) in the prostate tumor PC3 and LNCap cells (all P<0.05), and proliferation levels of the PC3 and LNCap cells significantly increased after the transfection (P<0.05). Conclusion: The CREB and p-CREB were highly expressed in human prostate cancer tissues and, the prostate cancer PC3 and LNCap cells cultured in vitro. In addition, the CREB and p-CREB could up-regulate the expressions of proliferation-related gene PCNA, and raise proliferation level of the cells.
2016, 23(2):230-234.
DOI: 10.3872/j.issn.1007-385X.2016.02.013
Abstract:
Objective:To investigate the expression of has miR-99b in gastric adenocarcinoma cell lines and its potential mechanism of affecting proliferation and migration of the gastric adenocarcinoma cell lines. Methods: Real-time PCR was performed to detect the expressions of miR-99b in human gastric epithelial cell line GES-1 and human gastric adenocarcinoma cell line (SGC7901, BGC823 and MGC-803). The synthesized miR-99b mimics were transfected into the SGC-7901 cells to constract a overexpressed cell line, and scramble mimics were transfected into the same cells as control group. Then proliferation and migration of the SGC-7901 cell lines which overexpressed miR-99b were measured by CCK-8 assay and transwell assay; expression level of mTOR (mammalian target of rapamycin) protein was determined by Western blotting. Results: The expression levels of miR-99b in the gastric adenocarcinoma cell lines, especially in the SGC-7901 cell line, were significantly lower than those in the human gastric epithelial cell line. After transfectio of the miR-99b mimics for 48 h, proliferation ability of the SGC-7901 cell in experiment group was lower than that of control group（0.20±0.00 vs 0.27±0.02，P<0.05）. Compared to the cell line transfected with the scramble mimics, the experiment group showed a significant reduction in number of transmembrane cells（226.27±25.17 vs 523.33±25.17，P<005）after the transfection for 24 h. miR-99b can downregulate the expression of mTOR protein in the human gastric adenocarcinoma cell line SGC-7901. Conclusion:The expression of miR-99b in gastric adenocarcinoma cell lines was obviously down regulated, inhibition effect of miR-99b on proliferation and migration of the human gastric adenocarcinoma cell line SGC-7901 might be related with down-regulated expression of mTOR.
Objective:To investigate the expression of has miR-99b in gastric adenocarcinoma cell lines and its potential mechanism of affecting proliferation and migration of the gastric adenocarcinoma cell lines. Methods: Real-time PCR was performed to detect the expressions of miR-99b in human gastric epithelial cell line GES-1 and human gastric adenocarcinoma cell line (SGC7901, BGC823 and MGC-803). The synthesized miR-99b mimics were transfected into the SGC-7901 cells to constract a overexpressed cell line, and scramble mimics were transfected into the same cells as control group. Then proliferation and migration of the SGC-7901 cell lines which overexpressed miR-99b were measured by CCK-8 assay and transwell assay; expression level of mTOR (mammalian target of rapamycin) protein was determined by Western blotting. Results: The expression levels of miR-99b in the gastric adenocarcinoma cell lines, especially in the SGC-7901 cell line, were significantly lower than those in the human gastric epithelial cell line. After transfectio of the miR-99b mimics for 48 h, proliferation ability of the SGC-7901 cell in experiment group was lower than that of control group（0.20±0.00 vs 0.27±0.02，P<0.05）. Compared to the cell line transfected with the scramble mimics, the experiment group showed a significant reduction in number of transmembrane cells（226.27±25.17 vs 523.33±25.17，P<005）after the transfection for 24 h. miR-99b can downregulate the expression of mTOR protein in the human gastric adenocarcinoma cell line SGC-7901. Conclusion:The expression of miR-99b in gastric adenocarcinoma cell lines was obviously down regulated, inhibition effect of miR-99b on proliferation and migration of the human gastric adenocarcinoma cell line SGC-7901 might be related with down-regulated expression of mTOR.
2016, 23(2):235-242.
DOI: 10.3872/j.issn.1007-385X.2016.02.014
Abstract:
Objective:To confirm the effect of ST3 β-galactosideα-2,3 sialyltransferase 5 (ST3GAL5) on drug resistance of AML in vivo and in vitro through studying the differential expression of ST3GAL5 between acute myeloid leukemia (AML) NB4 cell line and its drug-resistant cell line NB4/ADR. Methods: The expressions of ST3GAL5 in AML cell lines were detected by Real-time PCR and Western blotting assays. Expressions of ST3GAL5 in the cells were specifically regulated to increase or decrease. Before and after the interferences, Changes of sensitivities to chemotherapy drugs and activation of PI3K/Akt signaling pathway in the NB4 and NB4/ADR cells in vivo and in vitro were examined by MTT assay and experiments with xenograft mouse model. Results: ST3GAL5 was highly expressed in NB4 cell line but low expressed in its drug-resistant cell line NB4/ADR. Drug sensitivity and PI3K/Akt signaling pathway molecules of NB4/ADR cells in which expressions of ST3GAL5 were specifically up-regulated respectively increased and decreased (all P<0.05). Contrarily, drug sensitivity, PI3K/Akt signaling pathway molecules and P-gp of NB4 cells in which expressions of ST3GAL5 were down-regulated respectively decreased and increased. Conclusion: There were obvious differences of ST3GAL5 expressions between AML cell and its drug-resistant cell lines. These characteristic alterations are associated with multidrug resistance in AML, which is probably achieved through ST3GAL5 mediated-PI3K/Akt signaling pathway and alteration of P-gp expression.
Objective:To confirm the effect of ST3 β-galactosideα-2,3 sialyltransferase 5 (ST3GAL5) on drug resistance of AML in vivo and in vitro through studying the differential expression of ST3GAL5 between acute myeloid leukemia (AML) NB4 cell line and its drug-resistant cell line NB4/ADR. Methods: The expressions of ST3GAL5 in AML cell lines were detected by Real-time PCR and Western blotting assays. Expressions of ST3GAL5 in the cells were specifically regulated to increase or decrease. Before and after the interferences, Changes of sensitivities to chemotherapy drugs and activation of PI3K/Akt signaling pathway in the NB4 and NB4/ADR cells in vivo and in vitro were examined by MTT assay and experiments with xenograft mouse model. Results: ST3GAL5 was highly expressed in NB4 cell line but low expressed in its drug-resistant cell line NB4/ADR. Drug sensitivity and PI3K/Akt signaling pathway molecules of NB4/ADR cells in which expressions of ST3GAL5 were specifically up-regulated respectively increased and decreased (all P<0.05). Contrarily, drug sensitivity, PI3K/Akt signaling pathway molecules and P-gp of NB4 cells in which expressions of ST3GAL5 were down-regulated respectively decreased and increased. Conclusion: There were obvious differences of ST3GAL5 expressions between AML cell and its drug-resistant cell lines. These characteristic alterations are associated with multidrug resistance in AML, which is probably achieved through ST3GAL5 mediated-PI3K/Akt signaling pathway and alteration of P-gp expression.
2016, 23(2):243-249.
DOI: 10.3872/j.issn.1007-385X.2016.02.015
Abstract:
Objective:To study the effect of silencing CDK2 gene induced by the recombinant lentiviral vector pUL-CDK2-shRNA 3 (CDK2-shRNA) on the malignant biological behavior of mouse melanoma B16-F1 cells, and preliminarily investigate its mechanism.Methods: Melanoma B16-F1 cells were transfected by recombinant lentivirus vector CDK2-shRNA, and effect of which on proliferation of the B16-F1 cells was examined by MTT assay. Apoptosis of the cells was detected by DAPI staining and AnnexinV-FITC/ PI staining flow cytometry assay. Cell cycles were evaluated with flow cytometry assay. Cell adhesion assay and transwell chamber experiment were used to detect the changes of adhesion, invasion and migration abilities of the cells, respectively. Expression levels of proteins RB and pRB on signaling pathway, cell cycle related transcription factor E2F1 and migration related protein MMP-2 and MMP-9 were detected with Western blotting assay. Results: Compared with the blank control and negative control groups, relative proliferation and adhesion rates, migration and invasion abilities of the cells in the CDK2-shRNA group significantly decreased (P<0.05); while the apoptosis rate significantly increased (P<0.05); Number of the cells at G1 phase significantly increased, but number of the cells at S and G2 phases significantly decreased (P<0.05); expression of pRB and E2F1 proteins significantly decreased, but expression of RB significantly increased (P<0.05), which were related with cell cycles; and expression of MMP-2 and MMP-9 proteins for invasion and migration of the cells significantly decreased (P<0.05). Conclusions: Silencing CDK2 gene mediated by recombinant lentiviral vector pUL-CDK2-shRNA could significantly inhibit malignant biological behavior of the B16-F1 cells, and its mechanism might associate with the expression changes of the proteins for cell cycles, invasion and migration of the cells.
Objective:To study the effect of silencing CDK2 gene induced by the recombinant lentiviral vector pUL-CDK2-shRNA 3 (CDK2-shRNA) on the malignant biological behavior of mouse melanoma B16-F1 cells, and preliminarily investigate its mechanism.Methods: Melanoma B16-F1 cells were transfected by recombinant lentivirus vector CDK2-shRNA, and effect of which on proliferation of the B16-F1 cells was examined by MTT assay. Apoptosis of the cells was detected by DAPI staining and AnnexinV-FITC/ PI staining flow cytometry assay. Cell cycles were evaluated with flow cytometry assay. Cell adhesion assay and transwell chamber experiment were used to detect the changes of adhesion, invasion and migration abilities of the cells, respectively. Expression levels of proteins RB and pRB on signaling pathway, cell cycle related transcription factor E2F1 and migration related protein MMP-2 and MMP-9 were detected with Western blotting assay. Results: Compared with the blank control and negative control groups, relative proliferation and adhesion rates, migration and invasion abilities of the cells in the CDK2-shRNA group significantly decreased (P<0.05); while the apoptosis rate significantly increased (P<0.05); Number of the cells at G1 phase significantly increased, but number of the cells at S and G2 phases significantly decreased (P<0.05); expression of pRB and E2F1 proteins significantly decreased, but expression of RB significantly increased (P<0.05), which were related with cell cycles; and expression of MMP-2 and MMP-9 proteins for invasion and migration of the cells significantly decreased (P<0.05). Conclusions: Silencing CDK2 gene mediated by recombinant lentiviral vector pUL-CDK2-shRNA could significantly inhibit malignant biological behavior of the B16-F1 cells, and its mechanism might associate with the expression changes of the proteins for cell cycles, invasion and migration of the cells.
2016, 23(2):250-254.
DOI: 10.3872/j.issn.1007-385X.2016.02.016
Abstract:
Objective:To explore the role of chemokine CXCL12 and its receptor CXCR4 (biologic axis CXCL12/CXCR4) in promoting proliferation and invasion of endometrial carcinoma cells through transduction signaling pathway of extracellular signal-regulated kinase (ERK). Methods:Ishikawa endometrial carcinoma cell line was treated with exogenous CXCL12. The phosphorylation of ERK1/2 and the expression of survivin protein at different time points were detected by Western blotting; and secretion level of MMP-2 in supernatant of cell culture was measured by ELISA. At the same time, effects of AMD3100 and PD98059 on phosphorylation level of ERK1/2, level of Survivin protein and secretion level of MMP-2 were analyzed. Results: After treatment with exogenous CXCL12, phosphorylation level of ERK1/2 was rapidly risen (t=0.887，P<0.01）as well as expressions of Survivin and MMP-2 proteins were boosted(t=0.861，P<0.01; t=0.297，P<0.01) in a time dependent manner. Both of PD98059 and AMD3100 could significantly inhibit the phosphorylation level of p-ERK after induction with exogenous CXCL12. Combined action of PD98059 and AMD3100 could completely inhibit phosphorylation of ERK, block the activation of ERK pathway, and down-regulate the expressions of Survivin and MMP-2 proteins. Conclusion: Biologic axis of CXCL12/CXCR4 could up-regulate the expressions of Survivin and MMP-2 proteins through activation of ERK pathway, 〖JP3〗thus inspire a series of biological effects in proliferation and invasion of the Ishikawa cell.
Objective:To explore the role of chemokine CXCL12 and its receptor CXCR4 (biologic axis CXCL12/CXCR4) in promoting proliferation and invasion of endometrial carcinoma cells through transduction signaling pathway of extracellular signal-regulated kinase (ERK). Methods:Ishikawa endometrial carcinoma cell line was treated with exogenous CXCL12. The phosphorylation of ERK1/2 and the expression of survivin protein at different time points were detected by Western blotting; and secretion level of MMP-2 in supernatant of cell culture was measured by ELISA. At the same time, effects of AMD3100 and PD98059 on phosphorylation level of ERK1/2, level of Survivin protein and secretion level of MMP-2 were analyzed. Results: After treatment with exogenous CXCL12, phosphorylation level of ERK1/2 was rapidly risen (t=0.887，P<0.01）as well as expressions of Survivin and MMP-2 proteins were boosted(t=0.861，P<0.01; t=0.297，P<0.01) in a time dependent manner. Both of PD98059 and AMD3100 could significantly inhibit the phosphorylation level of p-ERK after induction with exogenous CXCL12. Combined action of PD98059 and AMD3100 could completely inhibit phosphorylation of ERK, block the activation of ERK pathway, and down-regulate the expressions of Survivin and MMP-2 proteins. Conclusion: Biologic axis of CXCL12/CXCR4 could up-regulate the expressions of Survivin and MMP-2 proteins through activation of ERK pathway, 〖JP3〗thus inspire a series of biological effects in proliferation and invasion of the Ishikawa cell.
LI Yu
,
HAN Jiaoling
,
ZHANG Zhena
,
WANG Fei
,
LI Hong
,
HUANG Jianmin
,
ZHAO Xuan
,
YANG Li
,
HUANG Lan
,
ZHANG Yi
2016, 23(2):255-260.
DOI: 10.3872/j.issn.1007-385X.2016.02.017
Abstract:
Objective:To explore the effect of combined action of anti-CD3 and anti-CD28 monoclonal antibodies on the proliferations and functions of cytokine-induced killer (CIK) cell and its subsets. Methods: Twenty peripheral blood samples of the patients with tumors (6 cases of renal carcinoma, 5 cases of lung cancer, every 2 cases of hepatic carcinoma, breast cancer and melanoma, and every 1 case of gallbladder carcinoma, lymphoma and ovarian cancer) who received biotherapy at the First Hospital Affiliated to Zhengzhou University from June 2014 to June 2015 were collected. Each sample of the peripheral blood was divided into four groups on average, namely control group, anti-CD3 monoclonal antibody alone group, anti-CD28 monoclonal antibody alone group and anti-CD3 combined with anti-CD28 monoclonal antibodies group, which were cultured with CIKs. Proliferation of the CIK cells was detected with CFSE staining. Secretion levels of IFN-γ, TNF-α and IL-2 by the CIK cells were detected by ELISA. CD4+, CD8+ and CD56+ cells of CIK cell subsets were sorted with magnetic beads. Then their abilities of secreting IFN-γ, TNF-α and IL-2 were examined by ELISA assay. Results:No significant difference in the proliferation ability of the CIK cells was detected between the anti-CD3/anti-CD28 combined group (PI=59.35) and anti-CD3 alone group (PI=64.4). However, combination of anti-CD3 and anti-CD28 monoclonal antibodies could stimulate CIK cells to secrete higher level of IFN-γ and TNF-α than those done by anti-CD3 alone, IFN-γ: (384.6±263.1) pg/ml vs (201.5±271.5)pg/ml, P=0.0361; TNF-α (4.795±1.251)pg/ml vs (2.835±0.443)pg/ml, P=0.0265. Furth analysis found that the combined action of anti-CD3 and anti-CD28 monoclonal antibodies could enhance activity of CD8+ cell subset to secret IFN-γ and activity of CD56+ cell subset to secret IL-2, thus strengthen the functions of lymphocytes. Conclusion: The combined action of anti-CD3 and anti-CD28 monoclonal antibodies could not enhance proliferation of the CIK cells, but strengthen function of the CIK cells to a certain extent, which has certain potential to be used in the immunotherapy of tumor cells.
Objective:To explore the effect of combined action of anti-CD3 and anti-CD28 monoclonal antibodies on the proliferations and functions of cytokine-induced killer (CIK) cell and its subsets. Methods: Twenty peripheral blood samples of the patients with tumors (6 cases of renal carcinoma, 5 cases of lung cancer, every 2 cases of hepatic carcinoma, breast cancer and melanoma, and every 1 case of gallbladder carcinoma, lymphoma and ovarian cancer) who received biotherapy at the First Hospital Affiliated to Zhengzhou University from June 2014 to June 2015 were collected. Each sample of the peripheral blood was divided into four groups on average, namely control group, anti-CD3 monoclonal antibody alone group, anti-CD28 monoclonal antibody alone group and anti-CD3 combined with anti-CD28 monoclonal antibodies group, which were cultured with CIKs. Proliferation of the CIK cells was detected with CFSE staining. Secretion levels of IFN-γ, TNF-α and IL-2 by the CIK cells were detected by ELISA. CD4+, CD8+ and CD56+ cells of CIK cell subsets were sorted with magnetic beads. Then their abilities of secreting IFN-γ, TNF-α and IL-2 were examined by ELISA assay. Results:No significant difference in the proliferation ability of the CIK cells was detected between the anti-CD3/anti-CD28 combined group (PI=59.35) and anti-CD3 alone group (PI=64.4). However, combination of anti-CD3 and anti-CD28 monoclonal antibodies could stimulate CIK cells to secrete higher level of IFN-γ and TNF-α than those done by anti-CD3 alone, IFN-γ: (384.6±263.1) pg/ml vs (201.5±271.5)pg/ml, P=0.0361; TNF-α (4.795±1.251)pg/ml vs (2.835±0.443)pg/ml, P=0.0265. Furth analysis found that the combined action of anti-CD3 and anti-CD28 monoclonal antibodies could enhance activity of CD8+ cell subset to secret IFN-γ and activity of CD56+ cell subset to secret IL-2, thus strengthen the functions of lymphocytes. Conclusion: The combined action of anti-CD3 and anti-CD28 monoclonal antibodies could not enhance proliferation of the CIK cells, but strengthen function of the CIK cells to a certain extent, which has certain potential to be used in the immunotherapy of tumor cells.
2016, 23(2):261-266.
DOI: 10.3872/j.issn.1007-385X.2016.02.018
Abstract:
Objective:To detect the methylation status of dishevelled-binding antagonist of beta-catenin 1 (DACT-1) gene in gastric cardia adenocarcinoma (GCA) tissues and adjacent non-cancerous tissues, and to study its clinical significance. Methods: Methylation specific PCR (MSP) and quantitative RT-PCR were respectively applied to examine the CpG methylation of the DACT-1 gene and expression of its mRNA in tumor tissues and corresponding adjacent noncancerous tissues of 112 samples (collected from Department of Surgery, the Fourth Hospital Affiliated to Hebei Medical University and Department of Thoracic Surgery, Cixian Cancer Hospital during 2006 to 2014). Results: Methylation rates of DACT-1 gene in the GCA tissues and the adjacent noncancerous tissues were 51.8% (58/112) and 17.6% (20/112) respectively, indicating that methylation rate of DACT-1 in the GCA tissues was significantly higher than that in the noncancerous tissues (P<0.01). Expression of DACT-1 mRNA in the cancer tissues was 0.580±0.143, which was significantly lower than that in the adjacent noncancerous tissues (0.654±0.110，P<0.01). Expression of DACT-1 mRNA in the GCA tissues with methylated DACT-1 was 0.488±0.097, which was obviously lower than that in the GCA tissues with unmetylated ACT-1 (0.675±0.120), and the methylation status of the gene was significantly related with expression of its mRNA（P<0.01). The hypermethylation of DACT-1 gene in the GCA tissues was significantly related with lymph node metastasis and family history of upper gastrointestinal cancer (P<0.05), but not related with age, gender, pathological grading and clinical staging of the cancer patients（P>0.05）. Conclusion: The hypermethylation of CpG in DACT-1 3gene might be one of the mechanisms causing down-regulation of DACT-1 expression. The methylation status of promoter area in DACT-1 gene is expected to provide a novel indicator for the clinical diagnosis and prognosis evaluation of GCA.
Objective:To detect the methylation status of dishevelled-binding antagonist of beta-catenin 1 (DACT-1) gene in gastric cardia adenocarcinoma (GCA) tissues and adjacent non-cancerous tissues, and to study its clinical significance. Methods: Methylation specific PCR (MSP) and quantitative RT-PCR were respectively applied to examine the CpG methylation of the DACT-1 gene and expression of its mRNA in tumor tissues and corresponding adjacent noncancerous tissues of 112 samples (collected from Department of Surgery, the Fourth Hospital Affiliated to Hebei Medical University and Department of Thoracic Surgery, Cixian Cancer Hospital during 2006 to 2014). Results: Methylation rates of DACT-1 gene in the GCA tissues and the adjacent noncancerous tissues were 51.8% (58/112) and 17.6% (20/112) respectively, indicating that methylation rate of DACT-1 in the GCA tissues was significantly higher than that in the noncancerous tissues (P<0.01). Expression of DACT-1 mRNA in the cancer tissues was 0.580±0.143, which was significantly lower than that in the adjacent noncancerous tissues (0.654±0.110，P<0.01). Expression of DACT-1 mRNA in the GCA tissues with methylated DACT-1 was 0.488±0.097, which was obviously lower than that in the GCA tissues with unmetylated ACT-1 (0.675±0.120), and the methylation status of the gene was significantly related with expression of its mRNA（P<0.01). The hypermethylation of DACT-1 gene in the GCA tissues was significantly related with lymph node metastasis and family history of upper gastrointestinal cancer (P<0.05), but not related with age, gender, pathological grading and clinical staging of the cancer patients（P>0.05）. Conclusion: The hypermethylation of CpG in DACT-1 3gene might be one of the mechanisms causing down-regulation of DACT-1 expression. The methylation status of promoter area in DACT-1 gene is expected to provide a novel indicator for the clinical diagnosis and prognosis evaluation of GCA.
2016, 23(2):267-275.
DOI: 10.3872/j.issn.1007-385X.2016.02.019
Abstract:
Objective:Value of high-expression of miR-17-92 cluster in evaluating prognosis of cancer patients was analyzed with Meta-analysis. Methods: English literatures on the relationship of high-expression of miR-17-92 cluster and the prognosis of cancer patients were retrieved from Web of Science, PubMed and EMBASE from construction of the data bases to Oct 1, 2015. Key data were extracted from the literatures, pooled hazard ratios (HRs) and corresponding 95% confidence interval (CI) were calculated. Results: Total of thirty-nine literatures was included, involving 4 908 patients. The high-expression of miR-17-92 cluster was associated with low overall survival rate of the patients with malignant tumor (HR=1.83, 95% CI: 1.58~2.12, P=0.000). Furthermore, disease-free survival rate (HR=1.80, 95% CI: 1.43~2.26, P=0.000), progression-free survival rate (HR=1.83, 95%CI: 1.11~3.02, P=0.018) and cancer-specific survival rate (HR=1.59, 95%CI: 1.04~2.42, P=0.032) in the cancer patients with high-expression of miR-17-92 cluster were all reduced. However, the pooled HRs for relapse- free survival rate was not statistically significant (P=0539). Conclusion: The high-expression of miR-17-92 cluster was associated with poor prognosis of the patients with malignant tumor, and it could be expected as a new marker for prognosis of the cancer patients.
Objective:Value of high-expression of miR-17-92 cluster in evaluating prognosis of cancer patients was analyzed with Meta-analysis. Methods: English literatures on the relationship of high-expression of miR-17-92 cluster and the prognosis of cancer patients were retrieved from Web of Science, PubMed and EMBASE from construction of the data bases to Oct 1, 2015. Key data were extracted from the literatures, pooled hazard ratios (HRs) and corresponding 95% confidence interval (CI) were calculated. Results: Total of thirty-nine literatures was included, involving 4 908 patients. The high-expression of miR-17-92 cluster was associated with low overall survival rate of the patients with malignant tumor (HR=1.83, 95% CI: 1.58~2.12, P=0.000). Furthermore, disease-free survival rate (HR=1.80, 95% CI: 1.43~2.26, P=0.000), progression-free survival rate (HR=1.83, 95%CI: 1.11~3.02, P=0.018) and cancer-specific survival rate (HR=1.59, 95%CI: 1.04~2.42, P=0.032) in the cancer patients with high-expression of miR-17-92 cluster were all reduced. However, the pooled HRs for relapse- free survival rate was not statistically significant (P=0539). Conclusion: The high-expression of miR-17-92 cluster was associated with poor prognosis of the patients with malignant tumor, and it could be expected as a new marker for prognosis of the cancer patients.
2016, 23(2):276-281.
DOI: 10.3872/j.issn.1007-385X.2016.02.020
Abstract:
Objective:To investigate the expressions of indoleamine 2,3-dioxygenase (IDO) and brdging integrator 1 (BIN1) in tumor tissues and tumor draining lymph nodes (TDLNs) of the patients with esophageal squamous cell carcinoma (ESCC) and their clinical significances. Methods: Pathological confirmed tumor tissues, carcinoma adjacent tissues and TDLNs were collected from 71 patients with ESCC who underwent tumor resection in the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University between Apr. 2013 and Jul. 2014. The carcinoma adjacent tissues were set up as the control group for tumor tissue group, while the non-metastatic lymph nodes set up as the control group for metastatic lymph node group. The expression levels of IDO and BIN1 were detected with Real-time PCR and immunohistochemistry assays, and correlation between the two expressions and their relations with clinical characteristics of the patients were analyzed. Results: Compared to the non-metastatic group, the expression level of IDO mRNA and positive rate of IDO protein were remarkably higher (0.47±0.14 vs 0.22±0.09, P<0.01; 90.91% vs 67.35%, P=0.042), while the expression level of BIN1 mRNA and positive rate of BIN1 protein remarkably lower (0.15±0.11 vs 0.35±015, P<0.01; 50.00% vs 77.55%, P=0.028) in metastatic group. In addition, the expression level of IDO mRNA and positive rate of IDO protein in carcinoma tissues were significantly higher than those in para-carcinoma tissues (0.51±0.12 vs 024±0.11, P<0.01; 81.69% vs 22.54%, P<0.01) while the expression level of BIN1 mRNA and positive rate of BIN1 protein significantly lower than those in para-carcinoma tissues (0.17±0.10 vs 0.41±0.14, P<001; 19.72% vs 80.28%, P=0.006). In both tumor tissues and TDLNs, the expression of IDO protein was associated with invasion range , lymph node metastasis and clinical stage of the tumors, while the expression of BIN1 protein was associated with degree of differentiation, invasion range, lymph node metastasis and clinical stage of the tumors. Conclusion: In ESCC tumor tissues and metastatic TDLNs, the expression level of IDO was significantly up-regulated and the expression level of BIN1 significantly down-regulated, which were closely associated with clinical features of the patients and might be important factors affecting progression of ESCC.
Objective:To investigate the expressions of indoleamine 2,3-dioxygenase (IDO) and brdging integrator 1 (BIN1) in tumor tissues and tumor draining lymph nodes (TDLNs) of the patients with esophageal squamous cell carcinoma (ESCC) and their clinical significances. Methods: Pathological confirmed tumor tissues, carcinoma adjacent tissues and TDLNs were collected from 71 patients with ESCC who underwent tumor resection in the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University between Apr. 2013 and Jul. 2014. The carcinoma adjacent tissues were set up as the control group for tumor tissue group, while the non-metastatic lymph nodes set up as the control group for metastatic lymph node group. The expression levels of IDO and BIN1 were detected with Real-time PCR and immunohistochemistry assays, and correlation between the two expressions and their relations with clinical characteristics of the patients were analyzed. Results: Compared to the non-metastatic group, the expression level of IDO mRNA and positive rate of IDO protein were remarkably higher (0.47±0.14 vs 0.22±0.09, P<0.01; 90.91% vs 67.35%, P=0.042), while the expression level of BIN1 mRNA and positive rate of BIN1 protein remarkably lower (0.15±0.11 vs 0.35±015, P<0.01; 50.00% vs 77.55%, P=0.028) in metastatic group. In addition, the expression level of IDO mRNA and positive rate of IDO protein in carcinoma tissues were significantly higher than those in para-carcinoma tissues (0.51±0.12 vs 024±0.11, P<0.01; 81.69% vs 22.54%, P<0.01) while the expression level of BIN1 mRNA and positive rate of BIN1 protein significantly lower than those in para-carcinoma tissues (0.17±0.10 vs 0.41±0.14, P<001; 19.72% vs 80.28%, P=0.006). In both tumor tissues and TDLNs, the expression of IDO protein was associated with invasion range , lymph node metastasis and clinical stage of the tumors, while the expression of BIN1 protein was associated with degree of differentiation, invasion range, lymph node metastasis and clinical stage of the tumors. Conclusion: In ESCC tumor tissues and metastatic TDLNs, the expression level of IDO was significantly up-regulated and the expression level of BIN1 significantly down-regulated, which were closely associated with clinical features of the patients and might be important factors affecting progression of ESCC.
2016, 23(2):282-286.
DOI: 10.3872/j.issn.1007-385X.2016.02.021
Abstract:
Notch信号通路是一种在进化中较为保守的信号通路，在多种生命活动中发挥重要作用。目前研究认为，Notch信号参与了免疫细胞的生长、分化和发育等多个环节的调控，其在B细胞的分化发育过程以及相关肿瘤的发生发展中起到重要的调控作用。本文对Notch信号在B细胞的不同分化发育阶段如淋巴祖细胞（common lymphoid progenitor，CLP）、边缘区B细胞（marginal zone B cell，MZ B）、滤泡B细胞（follicular B cell，FO B）、B-1 B细胞，以及B细胞淋巴瘤中的作用进行了综述。
Notch信号通路是一种在进化中较为保守的信号通路，在多种生命活动中发挥重要作用。目前研究认为，Notch信号参与了免疫细胞的生长、分化和发育等多个环节的调控，其在B细胞的分化发育过程以及相关肿瘤的发生发展中起到重要的调控作用。本文对Notch信号在B细胞的不同分化发育阶段如淋巴祖细胞（common lymphoid progenitor，CLP）、边缘区B细胞（marginal zone B cell，MZ B）、滤泡B细胞（follicular B cell，FO B）、B-1 B细胞，以及B细胞淋巴瘤中的作用进行了综述。
2016, 23(2):287-290.
DOI: 10.3872/j.issn.1007-385X.2016.02.022
Abstract:
T细胞基因转导方法的快速发展为肿瘤过继免疫治疗插上了飞翔的翅膀，近年来取得的颠覆性治疗效果让人们对它充满期待。T细胞的基因转导具有挑战性，以逆转录病毒、慢病毒、转座子和mRNA电穿孔为代表的技术进步，释放了T细胞的治疗潜能，实现了对肿瘤细胞精准、高效、持久地杀伤，大大提升了临床级肿瘤特异性T细胞的临床效果；但仍面临插入突变、转导效率、成本等方面的问题。相信随着相关基础研究的深入，T细胞基因转导“完美载体”的出现将会助力免疫治疗成为一线主流的肿瘤治疗方法。
T细胞基因转导方法的快速发展为肿瘤过继免疫治疗插上了飞翔的翅膀，近年来取得的颠覆性治疗效果让人们对它充满期待。T细胞的基因转导具有挑战性，以逆转录病毒、慢病毒、转座子和mRNA电穿孔为代表的技术进步，释放了T细胞的治疗潜能，实现了对肿瘤细胞精准、高效、持久地杀伤，大大提升了临床级肿瘤特异性T细胞的临床效果；但仍面临插入突变、转导效率、成本等方面的问题。相信随着相关基础研究的深入，T细胞基因转导“完美载体”的出现将会助力免疫治疗成为一线主流的肿瘤治疗方法。
2016, 23(2):291-296.
DOI: 10.3872/j.issn.1007-385X.2016.02.023
Abstract:
微小RNA（microRNA，miRNA）是目前肿瘤相关研究的热点，人类基因组中约有三分之一的基因受miRNAs调控。miR-370参与调节脂类代谢和肿瘤的发生、发展、耐药和转移，并发挥“癌基因”或“抑癌基因”的功能。越来越多的研究表明miR-370在人类多种肿瘤中异常表达，近年来也有研究认为脂类代谢异常是肿瘤形成的原因之一。本文就miR-370在肿瘤发生发展、复发转移和耐药中的作用及其作用机制进行综述。
微小RNA（microRNA，miRNA）是目前肿瘤相关研究的热点，人类基因组中约有三分之一的基因受miRNAs调控。miR-370参与调节脂类代谢和肿瘤的发生、发展、耐药和转移，并发挥“癌基因”或“抑癌基因”的功能。越来越多的研究表明miR-370在人类多种肿瘤中异常表达，近年来也有研究认为脂类代谢异常是肿瘤形成的原因之一。本文就miR-370在肿瘤发生发展、复发转移和耐药中的作用及其作用机制进行综述。
2016, 23(2):297-302.
DOI: 10.3872/j.issn.1007-385X.2016.02.024
Abstract:
长链非编码RNAs（long non-coding RNAs，lncRNAs）是一类长度超过200个核苷酸而无蛋白质编码功能的RNA分子，其在食管癌细胞的表观遗传学、转录及转录后水平参与基因的表达调控，在肿瘤的发生、发展、浸润和转移等过程中发挥致癌或抑癌基因的作用。本文就HOX转录反义RNA（HOX transcript antisense intergenic RNA，HOTAIR）、H19、肺腺癌转移相关转录子1（metastasis associated lung adenocarcinoma transcript l，MALAT1）等促癌相关lncRNAs，Epist等抑癌相关lncRNAs，边缘系统相关膜蛋白反义RNA3（limbicsystem-associated membrane protein antisense RNA3, LOC285194）及人类BOK反义RNA1（BOK antisense RNA 1，BOKAS1）等放化疗相关lncRNAs在食管癌中作用的研究进展作一综述。
长链非编码RNAs（long non-coding RNAs，lncRNAs）是一类长度超过200个核苷酸而无蛋白质编码功能的RNA分子，其在食管癌细胞的表观遗传学、转录及转录后水平参与基因的表达调控，在肿瘤的发生、发展、浸润和转移等过程中发挥致癌或抑癌基因的作用。本文就HOX转录反义RNA（HOX transcript antisense intergenic RNA，HOTAIR）、H19、肺腺癌转移相关转录子1（metastasis associated lung adenocarcinoma transcript l，MALAT1）等促癌相关lncRNAs，Epist等抑癌相关lncRNAs，边缘系统相关膜蛋白反义RNA3（limbicsystem-associated membrane protein antisense RNA3, LOC285194）及人类BOK反义RNA1（BOK antisense RNA 1，BOKAS1）等放化疗相关lncRNAs在食管癌中作用的研究进展作一综述。
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.