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Volume 22,Issue 6,2015 Table of Contents
2015, 22(6):675-683.
DOI: 10.3872/j.issn.1007-385X.2015.06.001
Abstract:
There are no standard effective systemic therapies for patients with hepatocellular carcinoma diagnosed at advanced stage, who have poor prognosis. It has been shown that Multi-target tyrosine kinase inhibitors (MTKIs) and adoptive NK cell immunotherapy have synergistic effect on hepatocellular carcinoma cells. In addition to blocking cell proliferation and angiogenesis signal pathway in tumor tissue to promote apoptosis, MTKIs also induce the expression of natural killer group 2 member D ligands (NKG2DLs) on tumor cells, which interact with NKG2D on NK cells to activate their antitumor activity. It is evident that MTKIs act on signaling molecules involved in DNA damage response, leading to activation of the alternative NF-κB complex that consists of NF-κB2 and RelB, which in turn increases the transcription of NKG2DLs. These results provided a mechanistic rationale for therapy using MTKIs together with adoptive NK cell transfer in the treatment of advanced hepatocellular carcinoma.
There are no standard effective systemic therapies for patients with hepatocellular carcinoma diagnosed at advanced stage, who have poor prognosis. It has been shown that Multi-target tyrosine kinase inhibitors (MTKIs) and adoptive NK cell immunotherapy have synergistic effect on hepatocellular carcinoma cells. In addition to blocking cell proliferation and angiogenesis signal pathway in tumor tissue to promote apoptosis, MTKIs also induce the expression of natural killer group 2 member D ligands (NKG2DLs) on tumor cells, which interact with NKG2D on NK cells to activate their antitumor activity. It is evident that MTKIs act on signaling molecules involved in DNA damage response, leading to activation of the alternative NF-κB complex that consists of NF-κB2 and RelB, which in turn increases the transcription of NKG2DLs. These results provided a mechanistic rationale for therapy using MTKIs together with adoptive NK cell transfer in the treatment of advanced hepatocellular carcinoma.
Huang Yuxian
,
Chen Xintong
,
Cai Songhao
,
Li Yuhua
,
Wu Bingyi
,
Song Chaoyang
,
He Yanjie
,
Guo Kunyuan
2015, 22(6):684-689.
DOI: 10.3872/j.issn.1007-385X.2015.06.002
Abstract:
Objective:To investigate the apoptotic effect of sunitinib on human hepatocellular carcinoma cell HepG2 and explore the underlying molecular mechanism. Methods: The HepG2 cells were cultivated by routine method. The effect of sunitinib on the growth of HepG2 cells was assessed by MTT assay. Molecular targets in the hepatocellular carcinoma HepG2 cells were examined by immunoblotting. Apoptotic cell death was detected using Annexin-V/PI double labeled flow cytometry and TUNEL assay. The expressions of mRNA were quantitated by RT-qPCR. Results: The IC50 of sunitinib for inhibiting HepG2 cell growth was （3.22±0.50）μmol/L. After exposed to sunitinib, the expression of VEGFR1, VEGFR2, PDGFRα, Kit, FLT3 were decreased in HepG2 cells. Apoptosis rates of HepG2 cells were (5.90±0.45)% vs (15.18±1.28)% in the absence or presence of sunitinib respectively, and corresponding apoptosis index (AI) were (4.17±0.64) vs (23.54±4.73). After treated with sunitinib, the expressions of pro-apoptotic genes Bax, NOXA, PUMA and P53 were increased in the cells, whereas that of apoptosis-inhibiting genes Bcl-2 and X-IAP were significantly decreased.Conclusion: Increased expression of pro-apoptotic genes and decreased apoptosis-inhibiting genes are likely responsible for sunitinib-induced apoptosis in human hepatocellular carcinoma HepG2 cells.
Objective:To investigate the apoptotic effect of sunitinib on human hepatocellular carcinoma cell HepG2 and explore the underlying molecular mechanism. Methods: The HepG2 cells were cultivated by routine method. The effect of sunitinib on the growth of HepG2 cells was assessed by MTT assay. Molecular targets in the hepatocellular carcinoma HepG2 cells were examined by immunoblotting. Apoptotic cell death was detected using Annexin-V/PI double labeled flow cytometry and TUNEL assay. The expressions of mRNA were quantitated by RT-qPCR. Results: The IC50 of sunitinib for inhibiting HepG2 cell growth was （3.22±0.50）μmol/L. After exposed to sunitinib, the expression of VEGFR1, VEGFR2, PDGFRα, Kit, FLT3 were decreased in HepG2 cells. Apoptosis rates of HepG2 cells were (5.90±0.45)% vs (15.18±1.28)% in the absence or presence of sunitinib respectively, and corresponding apoptosis index (AI) were (4.17±0.64) vs (23.54±4.73). After treated with sunitinib, the expressions of pro-apoptotic genes Bax, NOXA, PUMA and P53 were increased in the cells, whereas that of apoptosis-inhibiting genes Bcl-2 and X-IAP were significantly decreased.Conclusion: Increased expression of pro-apoptotic genes and decreased apoptosis-inhibiting genes are likely responsible for sunitinib-induced apoptosis in human hepatocellular carcinoma HepG2 cells.
Huang Yuxian
,
Chen Xintong
,
Lin Xia
,
Weng Guanyang
,
Li Yuhua
,
Wu Bingyi
,
Song Chaoyang
,
Guo Kunyuan
2015, 22(6):690-695.
DOI: 10.3872/j.issn.1007-385X.2015.06.003
Abstract:
Objective:To investigate the effect of sunitinib on natural killer group 2 member D ligands (NKG2DLs) expression and NK-mediated cytotoxicity in human hepatocellular carcinoma cells. Methods: HepG2 cells were cultivated by routine method. Human NK cells were isolated by magnetic activated cell sorting (MACS). Flow cytometry was used to evaluate the purity of the isolated NK cells and the expression levels of NKG2DLs on HepG2 cells. The cytotoxic effect of NK cells against HepG2 was assessed with LDH releasing assay. The expressions of NKG2DL mRNAs in HepG2 cells was quantitated by RT-qPCR. Results: More than 78% of the isolated cells were CD3-CD16+CD56+, indicative of NK cells. After sunitinib treatment, the expressions of multiple NKG2DLs on HepG2 cells were increased, especially that of MICA, MICB and ULBP2. At the E∶T ratio of 10∶1 and 20∶1, the cytotoxic effects of NK cells against HepG2 cells were increased from (9.47±1.11)% and (20.45±1.94)% in the untreated groups to (28.88±1.23)% and (44.93±157)% in the sunitinib treatment groups (P<0.05). The expression of MICA, MICB and ULBP2 mRNA in HepG2 cells was also significantly elevated after treated with sunitinib. Conclusion: Sunitinib regulates the expressions of NKG2DLs (MICA/B and ULBP2) on HepG2 cells, which activates NK cells and is responsible for their enhanced cytotoxic action.
Objective:To investigate the effect of sunitinib on natural killer group 2 member D ligands (NKG2DLs) expression and NK-mediated cytotoxicity in human hepatocellular carcinoma cells. Methods: HepG2 cells were cultivated by routine method. Human NK cells were isolated by magnetic activated cell sorting (MACS). Flow cytometry was used to evaluate the purity of the isolated NK cells and the expression levels of NKG2DLs on HepG2 cells. The cytotoxic effect of NK cells against HepG2 was assessed with LDH releasing assay. The expressions of NKG2DL mRNAs in HepG2 cells was quantitated by RT-qPCR. Results: More than 78% of the isolated cells were CD3-CD16+CD56+, indicative of NK cells. After sunitinib treatment, the expressions of multiple NKG2DLs on HepG2 cells were increased, especially that of MICA, MICB and ULBP2. At the E∶T ratio of 10∶1 and 20∶1, the cytotoxic effects of NK cells against HepG2 cells were increased from (9.47±1.11)% and (20.45±1.94)% in the untreated groups to (28.88±1.23)% and (44.93±157)% in the sunitinib treatment groups (P<0.05). The expression of MICA, MICB and ULBP2 mRNA in HepG2 cells was also significantly elevated after treated with sunitinib. Conclusion: Sunitinib regulates the expressions of NKG2DLs (MICA/B and ULBP2) on HepG2 cells, which activates NK cells and is responsible for their enhanced cytotoxic action.
Huang Yuxian
,
Chen Xintong
,
Cai Songhao
,
Long Hui
,
Guo Kunyuan
,
Wu Bingyi
,
Song Chaoyang
,
He Yanjie
,
Li Yuhua
2015, 22(6):696-702.
DOI: 10.3872/j.issn.1007-385X.2015.06.004
Abstract:
Objective: To investigate the interaction between the natural killer group 2 member D ligands (NKG2DLs) expressions and NF-κB signaling pathway in HepG2 cells treated with sunitinib.Methods: HepG2 cells were cultivated by routine method. The mRNAs of NF-κB gene family members in HepG2 cells were quantitated by RT-qPCR. The siRNAs specific for NF-κB1, NF-κB2 and RelB were designed using a computer soft and synthesized. They were transfected into HepG2 cells using liposome. The transfection and interference efficacy were evaluated by fluorescence microscopy, and real-time quantification PCR respectively. The expressions of NF-κB1, NF-κB2 and RelB were determined by immunoblotting, and the expressions of NKG2DLs were assessed by Flow cytometry.Results: The mRNAs of NF-κB1, and especially NF-κB2 and RelB were significantly increased in HepG2 cells treated with sunitinib. In siRNA transfection experiments, the red fluorescence was present in about 60% of HepG2 cells under fluorescence microscope, and the interference efficiency was 95%. Immunoblotting revealed that NF-κB1, NF-κB2 and RelB were decreased significantly in HepG2 cells after the transfection. Flow cytometry analysis found that the expressions of NKG2DLs in the siRNA transfected group was significantly lower than that in drug treatment group (P<005). Conclusion: Sunitinib induces the expressions of NKG2DLs on hepatocellular carcinoma HepG2 cells by the activating the alternative NF-κB pathway.
Objective: To investigate the interaction between the natural killer group 2 member D ligands (NKG2DLs) expressions and NF-κB signaling pathway in HepG2 cells treated with sunitinib.Methods: HepG2 cells were cultivated by routine method. The mRNAs of NF-κB gene family members in HepG2 cells were quantitated by RT-qPCR. The siRNAs specific for NF-κB1, NF-κB2 and RelB were designed using a computer soft and synthesized. They were transfected into HepG2 cells using liposome. The transfection and interference efficacy were evaluated by fluorescence microscopy, and real-time quantification PCR respectively. The expressions of NF-κB1, NF-κB2 and RelB were determined by immunoblotting, and the expressions of NKG2DLs were assessed by Flow cytometry.Results: The mRNAs of NF-κB1, and especially NF-κB2 and RelB were significantly increased in HepG2 cells treated with sunitinib. In siRNA transfection experiments, the red fluorescence was present in about 60% of HepG2 cells under fluorescence microscope, and the interference efficiency was 95%. Immunoblotting revealed that NF-κB1, NF-κB2 and RelB were decreased significantly in HepG2 cells after the transfection. Flow cytometry analysis found that the expressions of NKG2DLs in the siRNA transfected group was significantly lower than that in drug treatment group (P<005). Conclusion: Sunitinib induces the expressions of NKG2DLs on hepatocellular carcinoma HepG2 cells by the activating the alternative NF-κB pathway.
Huang Yuxian
,
Chen Xintong
,
Lin Xia
,
Weng Guanyang
,
Guo Kunyuan
,
Wu Bingyi
,
Song Chaoyang
,
He Yanjie
,
Li Yuhua
2015, 22(6):703-709.
DOI: 10.3872/j.issn.1007-385X.2015.06.005
Abstract:
Objective:To investigate molecular mechanism of sunitinib-induced expressions of natural killer group 2 member D ligands (NKG2DLs) in human hepatocellular carcinoma cell HepG2.Methods: HepG2 cells were cultivated by routine method. DNA damage in HepG2 cells was detected by single cell gel electrophoresis (SCGE) assay. The expressions of DNA damage-related molecule were quantitated by RT-qPCR. The levels of NKG2DLs, IKKα and IkBα were determined by immunoblotting. Results: Single cell gel electrophoresis revealed that HepG2 cells have various degree of DNA damages after exposed to sunitinib. RT-qPCR analysis showed that the expressions of AP-1, ATM, and ATR mRNA in HepG2 cells treated with sunitinib were significantly increased, whereas the levels of CHK1，CHK2，GSK3β mRNA were markedly lower. While the NF-κB inhibitor JSH-23 decreased the expressions of NKG2DLs in HepG2 cells, the agonist of NF-κB increased the expressions of these molecules. Furthermore, in HepG2 treated with sunitinib, IKKα was phosphorylated, and IkBα was activated. Conclusion: The data indicated that sunitinib induces the upregulated expressions of NKG2DLs in carcinoma cells by DNA damage-related molecules with activate the NF-κB pathway.
Objective:To investigate molecular mechanism of sunitinib-induced expressions of natural killer group 2 member D ligands (NKG2DLs) in human hepatocellular carcinoma cell HepG2.Methods: HepG2 cells were cultivated by routine method. DNA damage in HepG2 cells was detected by single cell gel electrophoresis (SCGE) assay. The expressions of DNA damage-related molecule were quantitated by RT-qPCR. The levels of NKG2DLs, IKKα and IkBα were determined by immunoblotting. Results: Single cell gel electrophoresis revealed that HepG2 cells have various degree of DNA damages after exposed to sunitinib. RT-qPCR analysis showed that the expressions of AP-1, ATM, and ATR mRNA in HepG2 cells treated with sunitinib were significantly increased, whereas the levels of CHK1，CHK2，GSK3β mRNA were markedly lower. While the NF-κB inhibitor JSH-23 decreased the expressions of NKG2DLs in HepG2 cells, the agonist of NF-κB increased the expressions of these molecules. Furthermore, in HepG2 treated with sunitinib, IKKα was phosphorylated, and IkBα was activated. Conclusion: The data indicated that sunitinib induces the upregulated expressions of NKG2DLs in carcinoma cells by DNA damage-related molecules with activate the NF-κB pathway.
2015, 22(6):710-715.
DOI: 10.3872/j.issn.1007-385X.2015.06.006
Abstract:
Objective:To investigate the cytotoxic effect of cytotoxic lymphocytes (CTLs) activated by Tat49-57-NY-ESO-1155-163 sensitized dendritic cells on melanoma A-375 cells. Methods:We collected the peripheral blood (about 50 ml) from healthy volunteers and isolated mononuclear cells by using lymphocyte separation medium. The cells were treated with cytokines to produce dendritic cells and T lymphocytes. After sensitized with the Tat49-57-NY-ESO-1155-163 peptide, the dendritic cells were co-cultured with the T lymphocyte cells to generate antigen-specific CTLs. Phenotypes of the dendritic cells were examined by flow cytometry. The antigen-specific cytotoxic activity of the CTLs against A-375 melanoma cells in vitro was assessed by the lactate dehydrogenase (LDH) method. Human lung cancer A549 cells and leukemia K562 cells were used as controls. Results: The expression rate of CD80/CD86 in dendritic cells sensitized with Tat49-57-NY-ESO-1155-163 was (54.9±3.3)% significantly higher than these sensitized with NY-ESO-1155-163 (\[43.8±5.7\]%，P<0.05). There were also significant increase of CD40 expression (\[42.1±1.9\]% vs \[23.7±2.8\]%，P<0.05) in Tat49-57-NY-ESO-1155-163 sensitized dendritic cells. These results indicated that the Tat fragment (49-57) significantly improved the cell penetrating ability of NY-ESO-1155-163. The T lymphocytes activated by the Tat49-57-NY-ESO-1155-163 sensitized DC were mainly CD3+CD8+cells. The cytotoxic activity of the CTLs induced by Tat49-57-NY-ESO-1155-163-sensitized dendritic cells toward A-375 melanoma cells was significantly higher than these induced with NY-ESO-1155-163-sensitized dendritic cells (P<0.05). The CTLs were also specific as they killed NY-ESO-1-expressing A375 more efficient than A549 cells and K562 cells (P<0.05). Conclusion: Tat49-57 cell penetrating peptide can enhance the immunogenicity of the peptide NY-ESO-1155-163. Tat49-57-NY-ESO-1155-163 polypeptide-sensitized dendritic cells can effectively induce the specific immune response against melanoma A-375 cells.
Objective:To investigate the cytotoxic effect of cytotoxic lymphocytes (CTLs) activated by Tat49-57-NY-ESO-1155-163 sensitized dendritic cells on melanoma A-375 cells. Methods:We collected the peripheral blood (about 50 ml) from healthy volunteers and isolated mononuclear cells by using lymphocyte separation medium. The cells were treated with cytokines to produce dendritic cells and T lymphocytes. After sensitized with the Tat49-57-NY-ESO-1155-163 peptide, the dendritic cells were co-cultured with the T lymphocyte cells to generate antigen-specific CTLs. Phenotypes of the dendritic cells were examined by flow cytometry. The antigen-specific cytotoxic activity of the CTLs against A-375 melanoma cells in vitro was assessed by the lactate dehydrogenase (LDH) method. Human lung cancer A549 cells and leukemia K562 cells were used as controls. Results: The expression rate of CD80/CD86 in dendritic cells sensitized with Tat49-57-NY-ESO-1155-163 was (54.9±3.3)% significantly higher than these sensitized with NY-ESO-1155-163 (\[43.8±5.7\]%，P<0.05). There were also significant increase of CD40 expression (\[42.1±1.9\]% vs \[23.7±2.8\]%，P<0.05) in Tat49-57-NY-ESO-1155-163 sensitized dendritic cells. These results indicated that the Tat fragment (49-57) significantly improved the cell penetrating ability of NY-ESO-1155-163. The T lymphocytes activated by the Tat49-57-NY-ESO-1155-163 sensitized DC were mainly CD3+CD8+cells. The cytotoxic activity of the CTLs induced by Tat49-57-NY-ESO-1155-163-sensitized dendritic cells toward A-375 melanoma cells was significantly higher than these induced with NY-ESO-1155-163-sensitized dendritic cells (P<0.05). The CTLs were also specific as they killed NY-ESO-1-expressing A375 more efficient than A549 cells and K562 cells (P<0.05). Conclusion: Tat49-57 cell penetrating peptide can enhance the immunogenicity of the peptide NY-ESO-1155-163. Tat49-57-NY-ESO-1155-163 polypeptide-sensitized dendritic cells can effectively induce the specific immune response against melanoma A-375 cells.
2015, 22(6):716-723.
DOI: 10.3872/j.issn.1007-385X.2015.06.007
Abstract:
Objective:To investigate the influence of downregulating Med19(mediator 19) expression on the therapeutic effect of paclitaxel in breast cancer cells with wild type or mutated p53 . Methods： Lentiviruses encoding small RNA that interferes Med19 expression were generated to infect cells of breast cancer cell lines MCF-7( p53 wild type) and MDA-MB-231 ( p53 mutation type). They were divided into Med19 knock-down groups (KD group, infected with pGcscil-Med19-siRNA-GFP), empty vector groups (NC group, infected with pGcscil-Med19-NC-GFP), and control groups (CON group, non-infected). GFP expression visualized under fluorescence microscope was used as an indicator of the efficiency of lentiviral infection. After silience of Med19 the levels of P53, phosphorylated P53 and P21 in cells of the different groups were determined by immunoblotting. The proliferation of MCF-7 and MDA-MB-231 cells under various experimental conditions was measured by the MTT assay. Flow cytometry was used to examine cell cycle and apoptosis. Results: Judged by fluorescent protein expression, more than 90% of MCF-7 cells and MDA-MB-231 cells infected with the lentiviruses in both KD and NC groups. The levels of Med19 expression in MCF-7 and MDA-MB-231 cells of the KD group were significantly lower than those in both CON and NC groups (P<0.05). The expressions of P53, phosphorylated P53, and P21 were significantly upregulated in MCF-7 cells of the KD group (P<0.05). In the 0.01-50 μg/ml range, paclitaxel induced a dose-dependent growth inhibition of both MCF-7 and MDA-MB-231 cells. For MCF-7 cells, the degree of inhibition and the number of apoptotic cells in KD group were significantly higher than those in the both NC and CON groups (P＜0.05). However, they were not significantly different between the three groups of MDA-MB-231 cells (P>0.05). While both MCF-7 and MDA-MB-231 cells underwent G2/M cell cycle block when treated with 10 μg/ml of paclitaxel, cycle of G0/G1 phase in KD group of MCF-7 cells more significantly increased than that in their NC group (P<005). Conclusion: The therapeutic effect of paclitaxel on MCF-7 cells( p53wild type) is enhanced by silencing Med19, which is likely mediated by the activation of p53 and expression of its downstream genes including p21 gene, leading to cell cycleb lock and apoptosis.
Objective:To investigate the influence of downregulating Med19(mediator 19) expression on the therapeutic effect of paclitaxel in breast cancer cells with wild type or mutated p53 . Methods： Lentiviruses encoding small RNA that interferes Med19 expression were generated to infect cells of breast cancer cell lines MCF-7( p53 wild type) and MDA-MB-231 ( p53 mutation type). They were divided into Med19 knock-down groups (KD group, infected with pGcscil-Med19-siRNA-GFP), empty vector groups (NC group, infected with pGcscil-Med19-NC-GFP), and control groups (CON group, non-infected). GFP expression visualized under fluorescence microscope was used as an indicator of the efficiency of lentiviral infection. After silience of Med19 the levels of P53, phosphorylated P53 and P21 in cells of the different groups were determined by immunoblotting. The proliferation of MCF-7 and MDA-MB-231 cells under various experimental conditions was measured by the MTT assay. Flow cytometry was used to examine cell cycle and apoptosis. Results: Judged by fluorescent protein expression, more than 90% of MCF-7 cells and MDA-MB-231 cells infected with the lentiviruses in both KD and NC groups. The levels of Med19 expression in MCF-7 and MDA-MB-231 cells of the KD group were significantly lower than those in both CON and NC groups (P<0.05). The expressions of P53, phosphorylated P53, and P21 were significantly upregulated in MCF-7 cells of the KD group (P<0.05). In the 0.01-50 μg/ml range, paclitaxel induced a dose-dependent growth inhibition of both MCF-7 and MDA-MB-231 cells. For MCF-7 cells, the degree of inhibition and the number of apoptotic cells in KD group were significantly higher than those in the both NC and CON groups (P＜0.05). However, they were not significantly different between the three groups of MDA-MB-231 cells (P>0.05). While both MCF-7 and MDA-MB-231 cells underwent G2/M cell cycle block when treated with 10 μg/ml of paclitaxel, cycle of G0/G1 phase in KD group of MCF-7 cells more significantly increased than that in their NC group (P<005). Conclusion: The therapeutic effect of paclitaxel on MCF-7 cells( p53wild type) is enhanced by silencing Med19, which is likely mediated by the activation of p53 and expression of its downstream genes including p21 gene, leading to cell cycleb lock and apoptosis.
Li Xiuping
,
Xu Zhiwei
,
Liu Baojin
,
Wu Chongxue
,
Zhu Kunchao
,
Xie Liang
,
Zhu Xiaofeng
,
Su Weiwei
,
Li Haijia
,
Meng Xuli
2015, 22(6):724-628.
DOI: 10.3872/j.issn.1007-385X.2015.06.008
Abstract:
Objective:To investigate the potential mechanism by which tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) inhibits the invasion capability of breast cancer MDA-MB-231 cells. Methods：After treating MDA-MB-231 cells with rsTRAIL, the expression of EGFR and p-IκBα were examined by immunoblotting, and the level of miR-146 was measured by real-time quantitative PCR. Immunoblotting was also used to detect the effect of miR-146a on EGFR expression. Transwell assay was carried out to assess the effects of rsTRAIL, miR-146a and EGFR on the invasion ability of MDA-MB-231 cells. Results: In MDA-MB-231 cells treated for 6, 12, and 24 hours, rsTRAIL(20 ng/ml) significantly suppressed the expression of EGFR (6 h, t=4.35, P<0.05; 12 h, t=8.609, P<0.01; 24 h, t=1084, P<0.01), increased the level of the phosphorylated IκBα (6 h, t=-4.201, P<0.05; 12 h, t =-15.805, P<001; 24 h, t=-35.921, P<0.01), and upregulated the expression of miR-146a (6 h, t=-4.67,P<0.05; 12 h, t=-11.635, P<0.01;24 h, t=-15.8, P<0.01), and on time dependent. Compared with that in control cells, the level of EGFR (t=625, P<0.01) was significantly decreased in MDA-MB-231 cells transfected with miR-146a mimics, whereas in the same cells transfected with miR-146 inhibitor the expression of EGFR promoted (t=-3.674, P<005). Furthermore, transfection with rsTRAIL, as well as transfection with miR-146a mimics or siEGFRall dramatically decreased the invasion ability of MDA-MB-231 cells (t=7.108, P<0.01; t=6.051, P<0.01; t=5.245, P<0.01 respectively). Conclusion: rsTRAIL specifically suppresses EGFR-dependent invasion capability of human breast cancer through inducing increased expression of miR-146a.
Objective:To investigate the potential mechanism by which tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) inhibits the invasion capability of breast cancer MDA-MB-231 cells. Methods：After treating MDA-MB-231 cells with rsTRAIL, the expression of EGFR and p-IκBα were examined by immunoblotting, and the level of miR-146 was measured by real-time quantitative PCR. Immunoblotting was also used to detect the effect of miR-146a on EGFR expression. Transwell assay was carried out to assess the effects of rsTRAIL, miR-146a and EGFR on the invasion ability of MDA-MB-231 cells. Results: In MDA-MB-231 cells treated for 6, 12, and 24 hours, rsTRAIL(20 ng/ml) significantly suppressed the expression of EGFR (6 h, t=4.35, P<0.05; 12 h, t=8.609, P<0.01; 24 h, t=1084, P<0.01), increased the level of the phosphorylated IκBα (6 h, t=-4.201, P<0.05; 12 h, t =-15.805, P<001; 24 h, t=-35.921, P<0.01), and upregulated the expression of miR-146a (6 h, t=-4.67,P<0.05; 12 h, t=-11.635, P<0.01;24 h, t=-15.8, P<0.01), and on time dependent. Compared with that in control cells, the level of EGFR (t=625, P<0.01) was significantly decreased in MDA-MB-231 cells transfected with miR-146a mimics, whereas in the same cells transfected with miR-146 inhibitor the expression of EGFR promoted (t=-3.674, P<005). Furthermore, transfection with rsTRAIL, as well as transfection with miR-146a mimics or siEGFRall dramatically decreased the invasion ability of MDA-MB-231 cells (t=7.108, P<0.01; t=6.051, P<0.01; t=5.245, P<0.01 respectively). Conclusion: rsTRAIL specifically suppresses EGFR-dependent invasion capability of human breast cancer through inducing increased expression of miR-146a.
2015, 22(6):729-733.
DOI: 10.3872/j.issn.1007-385X.2015.06.009
Abstract:
Objective:To investigate the effect of Artemisinin on apoptosis of non-small cell lung cancer (NSCLC) ASTC-a-1 and A549 cells and its mechanism. Methods:CCK-8 assay was performed to assess the effect of treatment with Artemisinin at 0-150 μg/ml for 24 h and at 100 μg/ml for 0, 6, 12, 24, 48 h on the activity of ASTC-a-1 and A549 cells. Laser scanning confocal microscope was used to observed influence of treatment with STS at 1 μg/ml, Artemisinin at 100 μg/ml alone or combined with NAC for 24 h on nuclear morphologies, and flow cytometry was used to examine impact of treatment with Artemisinin alone or combined with NAC on apoptosis of the cells; After silencing of Bax and Bak genes by RNA interference technology, effect of Artemisinin on viability of the cells was examined.Results: Artemisinin induced growth inhibition of ASTC-a-1 and A549 cells in a concentration and time dependent manner (IC50≈100 μg/ml), and the falling level of viability of the cells induced by Artemisinin was significantly less than untreaded cells after pretreated with NAC (all P<0.01). Treatments of ASTC-a-1 and A549 cells with STS, Artemisinin alone or combined with NAC induced their karyopyknosis and apoptosis. After pretreatment with NAC, the apoptosis rates of ASTC-a-1 and A549 cells induced by Artemisinin were (20.4±2.1)% and (17.9±3.8)% respectively, which were obviously less than STS group ( \[48.2±2.6\]% and \[39.8±4.9\]%, respectively) and without pretreated group (\[59.6±3.4\]% and \[50.7±3.8\]% respectively) (all P<0.01). Artemisinin only increased viability of silencing Bak cells (P<0.01), but not obviously impact that of silencing Bax cells (P＞0.05). Conclusion: Artemisinin induces apoptosis of non-small cell lung cancer ASTC-a-1 and A549 cells, which requires the participation of reactive oxygen species (ROS) and promoting apoptosis factor Bak, but not Bax.
Objective:To investigate the effect of Artemisinin on apoptosis of non-small cell lung cancer (NSCLC) ASTC-a-1 and A549 cells and its mechanism. Methods:CCK-8 assay was performed to assess the effect of treatment with Artemisinin at 0-150 μg/ml for 24 h and at 100 μg/ml for 0, 6, 12, 24, 48 h on the activity of ASTC-a-1 and A549 cells. Laser scanning confocal microscope was used to observed influence of treatment with STS at 1 μg/ml, Artemisinin at 100 μg/ml alone or combined with NAC for 24 h on nuclear morphologies, and flow cytometry was used to examine impact of treatment with Artemisinin alone or combined with NAC on apoptosis of the cells; After silencing of Bax and Bak genes by RNA interference technology, effect of Artemisinin on viability of the cells was examined.Results: Artemisinin induced growth inhibition of ASTC-a-1 and A549 cells in a concentration and time dependent manner (IC50≈100 μg/ml), and the falling level of viability of the cells induced by Artemisinin was significantly less than untreaded cells after pretreated with NAC (all P<0.01). Treatments of ASTC-a-1 and A549 cells with STS, Artemisinin alone or combined with NAC induced their karyopyknosis and apoptosis. After pretreatment with NAC, the apoptosis rates of ASTC-a-1 and A549 cells induced by Artemisinin were (20.4±2.1)% and (17.9±3.8)% respectively, which were obviously less than STS group ( \[48.2±2.6\]% and \[39.8±4.9\]%, respectively) and without pretreated group (\[59.6±3.4\]% and \[50.7±3.8\]% respectively) (all P<0.01). Artemisinin only increased viability of silencing Bak cells (P<0.01), but not obviously impact that of silencing Bax cells (P＞0.05). Conclusion: Artemisinin induces apoptosis of non-small cell lung cancer ASTC-a-1 and A549 cells, which requires the participation of reactive oxygen species (ROS) and promoting apoptosis factor Bak, but not Bax.
2015, 22(6):734-739.
DOI: 10.3872/j.issn.1007-385X.2015.06.010
Abstract:
Objective:To provide guide for the clinical medication of EGFR-tyrosine kinase inhibitors (TKIs) and discussion of their association with clinical pathological features, we investigated the amplification and mutation status of genes encoding epidermal growth factor receptor (EGFR), Kirsten rat sarcoma viral oncogene homolog (KRAS), B-Raf proto oncogene serine/threonine protein kinase (BRAF) and phosphatidylinositol -4，5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) in NSCLC patients.Methods:EGFR, KRAS, BRAF and PIK3CA mutations in 430 randomly selected Chinese patients with NSCLC were analyzed by SurPlex-xTAG70plex platform. The relationship between the mutations and the clinicopathologic features was further evaluated.Results: The mutation rates of EGFR, KRAS, BRAF and PIK3CA were 41.2%, 7.9%, 0.7%, and 3.7% respectively in these patients. The mutation rates of EGFR exon 19 and 21 were higher in females than those in males (P<0.01), significantly increased in adenocarcinomas compared to those in the other forms of lung cancers (P<0.01), and risen markedly in non-smokers compared to those in smokers (P<001). Conversely, the KRAS mutation rates were higher in males than those in females (P<0.05), increased significantly in adenocarcinomas compared to those in the other forms of lung cancers (P<0.005), and risen markedly in smokers compared to those in non-smokers (P<0.01). The PIK3CA mutation rates were significantly lower in adenocarcinomas compared to those in the other forms of lung cancers (P<0.01). Conclusion: The mutation rates of EGFR and KRAS in NSCLC are associated with gender, pathohistology, and smoking habits. Concurrent presence of EGFR and KRAS mutations was found in NSCLC from these patients, and the mutational statuses of PIK3CA and EGFR or KRAS were not mutually exclusive.
Objective:To provide guide for the clinical medication of EGFR-tyrosine kinase inhibitors (TKIs) and discussion of their association with clinical pathological features, we investigated the amplification and mutation status of genes encoding epidermal growth factor receptor (EGFR), Kirsten rat sarcoma viral oncogene homolog (KRAS), B-Raf proto oncogene serine/threonine protein kinase (BRAF) and phosphatidylinositol -4，5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) in NSCLC patients.Methods:EGFR, KRAS, BRAF and PIK3CA mutations in 430 randomly selected Chinese patients with NSCLC were analyzed by SurPlex-xTAG70plex platform. The relationship between the mutations and the clinicopathologic features was further evaluated.Results: The mutation rates of EGFR, KRAS, BRAF and PIK3CA were 41.2%, 7.9%, 0.7%, and 3.7% respectively in these patients. The mutation rates of EGFR exon 19 and 21 were higher in females than those in males (P<0.01), significantly increased in adenocarcinomas compared to those in the other forms of lung cancers (P<0.01), and risen markedly in non-smokers compared to those in smokers (P<001). Conversely, the KRAS mutation rates were higher in males than those in females (P<0.05), increased significantly in adenocarcinomas compared to those in the other forms of lung cancers (P<0.005), and risen markedly in smokers compared to those in non-smokers (P<0.01). The PIK3CA mutation rates were significantly lower in adenocarcinomas compared to those in the other forms of lung cancers (P<0.01). Conclusion: The mutation rates of EGFR and KRAS in NSCLC are associated with gender, pathohistology, and smoking habits. Concurrent presence of EGFR and KRAS mutations was found in NSCLC from these patients, and the mutational statuses of PIK3CA and EGFR or KRAS were not mutually exclusive.
2015, 22(6):740-746.
DOI: 10.3872/j.issn.1007-385X.2015.06.011
Abstract:
Objective:To identify molecular markers for accurate diagnosis of thyroid cancer the molecular phenotypes of varied proteiumark were evaluated in differentiated thyroid cancer (DTC) and benign thyroid lesions. Methods：Tissue microarrays containing 100 benign and 99 malignant thyroid lesions were stained for a panel of 57 molecular markers. Correlation between the marker staining and tumor pathology (DTC versus benign tumor) were determined using contingency table and Mann-Whitney U (MU) tests. A Random Forests classifier algorithm was also utilized to identify meaningful important molecular classifiers. Results： Of the 57 diagnostic markers evaluated, 35 (61%) were significantly associated with a DTC diagnosis after multiple testing correction. Among them, 8 markers were downregulated and 27 ones upregulated in DTC, when compared with benign thyroid tumor. The most significant markers for DTC diagnosis were Galectin-3, Cytokeratin 19, Vascular Endothelial Growth Factor, Androgen Receptor, p16, Aurora-A, and HBME-1. Using the entire molecular marker panel, a Random Forests algorithm was able to classify a neoplasm as DTC or benign tumor with an estimated sensitivity of 87.9%, specificity of 94.0%, and accuracy of 91.0%. Conclusion：Evaluating the DTC and benign thyroid tumor molecular phenotypes has allowed us to identify a diagnostic marker panel, which may help differentiate DTC with benign tumors and improve patient selection for thyroid surgery.
Objective:To identify molecular markers for accurate diagnosis of thyroid cancer the molecular phenotypes of varied proteiumark were evaluated in differentiated thyroid cancer (DTC) and benign thyroid lesions. Methods：Tissue microarrays containing 100 benign and 99 malignant thyroid lesions were stained for a panel of 57 molecular markers. Correlation between the marker staining and tumor pathology (DTC versus benign tumor) were determined using contingency table and Mann-Whitney U (MU) tests. A Random Forests classifier algorithm was also utilized to identify meaningful important molecular classifiers. Results： Of the 57 diagnostic markers evaluated, 35 (61%) were significantly associated with a DTC diagnosis after multiple testing correction. Among them, 8 markers were downregulated and 27 ones upregulated in DTC, when compared with benign thyroid tumor. The most significant markers for DTC diagnosis were Galectin-3, Cytokeratin 19, Vascular Endothelial Growth Factor, Androgen Receptor, p16, Aurora-A, and HBME-1. Using the entire molecular marker panel, a Random Forests algorithm was able to classify a neoplasm as DTC or benign tumor with an estimated sensitivity of 87.9%, specificity of 94.0%, and accuracy of 91.0%. Conclusion：Evaluating the DTC and benign thyroid tumor molecular phenotypes has allowed us to identify a diagnostic marker panel, which may help differentiate DTC with benign tumors and improve patient selection for thyroid surgery.
2015, 22(6):747-753.
DOI: 10.3872/j.issn.1007-385X.2015.06.012
Abstract:
Objective:To investigate the expression of protein S6 kinase1（S6K1）and eukaryotic translation initiation factor 4E binding protein 1（4EBP1）in colorectal cancers and their relationship with clinicopathologic characteristics. Methods:The data of 100 patients diagnosed with colorectal cancer from January of 2007 to December of 2009 were collection from the General Hospital of Ji'nan Military Command of PLA. The specimens include 100 colorectal cancer tissues, 40 normal tissues of the incisional edges, 30 metastatic lymph nodes, and 15 distant metastasis cancer tissues. Immunohistochemical S-P method was used to examine the expression of S6K1 and 4EBP1 in these tissues. Results: The positive rates of S6K1 and 4EBP1 were 17.50% (7/40) and 22.50% (9/40) respectively in normal colorectal tissues, and they increased significantly to 74.00% (74/100) and 79.00% (79/100) in colorectal cancer tissues. The expression of 4EBP1 in colorectal cancer tissues was associated with its clinical stage and differentiation status (P<0.05). Patients with colorectal cancers expressing high levels of S6K1 and 4EBP1 had shorter progression free survival (PFS) (P<0.05), whereas S6K1 and 4EBP1 expression had no association with overall survival (OS) (P>0.05). Further analysis revealed that there was a direct correlation between the levels of S6K1 and 4EBP1 in colorectal cancer tissues (r=0.246, P<0.05). Conclusion:The expression of S6K1 and 4EBP1 are related to the development, progress, and prognosis of colorectal cancer. Therefore, they are potential therapeutic targets and informative prognostic biomarkers.
Objective:To investigate the expression of protein S6 kinase1（S6K1）and eukaryotic translation initiation factor 4E binding protein 1（4EBP1）in colorectal cancers and their relationship with clinicopathologic characteristics. Methods:The data of 100 patients diagnosed with colorectal cancer from January of 2007 to December of 2009 were collection from the General Hospital of Ji'nan Military Command of PLA. The specimens include 100 colorectal cancer tissues, 40 normal tissues of the incisional edges, 30 metastatic lymph nodes, and 15 distant metastasis cancer tissues. Immunohistochemical S-P method was used to examine the expression of S6K1 and 4EBP1 in these tissues. Results: The positive rates of S6K1 and 4EBP1 were 17.50% (7/40) and 22.50% (9/40) respectively in normal colorectal tissues, and they increased significantly to 74.00% (74/100) and 79.00% (79/100) in colorectal cancer tissues. The expression of 4EBP1 in colorectal cancer tissues was associated with its clinical stage and differentiation status (P<0.05). Patients with colorectal cancers expressing high levels of S6K1 and 4EBP1 had shorter progression free survival (PFS) (P<0.05), whereas S6K1 and 4EBP1 expression had no association with overall survival (OS) (P>0.05). Further analysis revealed that there was a direct correlation between the levels of S6K1 and 4EBP1 in colorectal cancer tissues (r=0.246, P<0.05). Conclusion:The expression of S6K1 and 4EBP1 are related to the development, progress, and prognosis of colorectal cancer. Therefore, they are potential therapeutic targets and informative prognostic biomarkers.
2015, 22(6):754-759.
DOI: 10.3872/j.issn.1007-385X.2015.06.013
Abstract:
Objective:To investigate the expression and methylation status of lncRNA XLOC_008370 gene in esophageal squamous cell carcinoma（ESCC）and explore their role in ESCC development. Methods: RT-PCR and methylation specific PCR (MSP) were used to examine the expression and methylation status of XLOC_008370 gene in esophageal cancer cell lines (TE1, TE13, T.TN, Eca109, Yes-2) and in primary ESCC tissues. Their relationships with clinical pathological feature were further analyzed.Results: All 5 lines of esophageal cancer cells express no or low level of XLOC_008370, which was increased significantly after exposed to 5-Aza-dC, a DNA methyltransferase inhibitor. While the XLOC_008370 gene was highly methylated in all the 5 lines of cells, treatment with 5-Aza-dC led to decreased methylation of the gene in the Eca109 and Yes-2 cells, and completely demethylation in the TE1, TE13, and T.TN cells. In primary ESCC tumor tissues, the level of XLOC_008370 was significantly lower compared to that in corresponding non-cancerous tissues (P<0.05), and the expression was associated with TNM and pathological stages (P<0.05). The methylation frequency of XLOC_008370 gene promoter in ESCC tissues (54.02%, 47/87) was significantly higher than that in corresponding normal tissues (9.20%, 8/87) (P<0.05), and it was also associated with TNM and pathological stages (P<0.05). Furthermore, the level of XLOC_008370 in ESCC tumor tissues having its gene methylated was significantly lower than that in tumor tissues having the gene unmethylated (P<0.05). Conclusion: 〗Aberrant low expression of lncRNA XLOC_008370 is closely related to the development of ESCC, and promoter methylation is likely one of the mechanisms responsible for its decreased expression.
Objective:To investigate the expression and methylation status of lncRNA XLOC_008370 gene in esophageal squamous cell carcinoma（ESCC）and explore their role in ESCC development. Methods: RT-PCR and methylation specific PCR (MSP) were used to examine the expression and methylation status of XLOC_008370 gene in esophageal cancer cell lines (TE1, TE13, T.TN, Eca109, Yes-2) and in primary ESCC tissues. Their relationships with clinical pathological feature were further analyzed.Results: All 5 lines of esophageal cancer cells express no or low level of XLOC_008370, which was increased significantly after exposed to 5-Aza-dC, a DNA methyltransferase inhibitor. While the XLOC_008370 gene was highly methylated in all the 5 lines of cells, treatment with 5-Aza-dC led to decreased methylation of the gene in the Eca109 and Yes-2 cells, and completely demethylation in the TE1, TE13, and T.TN cells. In primary ESCC tumor tissues, the level of XLOC_008370 was significantly lower compared to that in corresponding non-cancerous tissues (P<0.05), and the expression was associated with TNM and pathological stages (P<0.05). The methylation frequency of XLOC_008370 gene promoter in ESCC tissues (54.02%, 47/87) was significantly higher than that in corresponding normal tissues (9.20%, 8/87) (P<0.05), and it was also associated with TNM and pathological stages (P<0.05). Furthermore, the level of XLOC_008370 in ESCC tumor tissues having its gene methylated was significantly lower than that in tumor tissues having the gene unmethylated (P<0.05). Conclusion: 〗Aberrant low expression of lncRNA XLOC_008370 is closely related to the development of ESCC, and promoter methylation is likely one of the mechanisms responsible for its decreased expression.
2015, 22(6):760-764.
DOI: 10.3872/j.issn.1007-385X.2015.06.014
Abstract:
Objective: To assess the clinical efficacy and safety of the combined therapy with oxaliplatin (OXA)-based regimen and DC-CIK (dendritic cells and cytokine-induced killer) cells for the treatment of advanced primary liver carcinoma.Methods：A retrospective analysis was carried out for 11 patients with advanced primary liver carcinoma enrolled into the Department of Biotherapy of Eastern Hepatobiliary Surgery Hospital from Jan. 2013 to Jun. 2015. All these patients received FOLFOX4 (OXA+calcium folinatc \[CF\]+5-fluorouracil\[5-FU\]）or GEMOX (gemcitabine\[GEM\]+OXA) chemotherapy. After 2-3 days of chemotherapy, DC-CIK cells immune therapy was administrated. Disease control rate(DCR), median time to tumor progression (mTTP), median overall survival (mOS), and adverse effects were recorded. Results：Among the 11 patients，no one had CR，2 had PR, 5 had SD, and 4 had PD. The response rate and disease control rate were 2/11 and 7/11 respectively. The mTTP and mOS were 4.1 months and 11.3 months. The major side effects were mild myelosuppression，gastrointestinal reaction and peripheral nerve toxicity, which were manageable with symptomatic treatments. Conclusion: For the treatment of advanced primary liver carcinoma, the combined therapy with oxaliplatin-based regimen and DC-CIK cells is effective, and its adverse effects are mild and well-tolerated, indicating that the therapy deserves further evaluation in more clinical trials with more patients.
Objective: To assess the clinical efficacy and safety of the combined therapy with oxaliplatin (OXA)-based regimen and DC-CIK (dendritic cells and cytokine-induced killer) cells for the treatment of advanced primary liver carcinoma.Methods：A retrospective analysis was carried out for 11 patients with advanced primary liver carcinoma enrolled into the Department of Biotherapy of Eastern Hepatobiliary Surgery Hospital from Jan. 2013 to Jun. 2015. All these patients received FOLFOX4 (OXA+calcium folinatc \[CF\]+5-fluorouracil\[5-FU\]）or GEMOX (gemcitabine\[GEM\]+OXA) chemotherapy. After 2-3 days of chemotherapy, DC-CIK cells immune therapy was administrated. Disease control rate(DCR), median time to tumor progression (mTTP), median overall survival (mOS), and adverse effects were recorded. Results：Among the 11 patients，no one had CR，2 had PR, 5 had SD, and 4 had PD. The response rate and disease control rate were 2/11 and 7/11 respectively. The mTTP and mOS were 4.1 months and 11.3 months. The major side effects were mild myelosuppression，gastrointestinal reaction and peripheral nerve toxicity, which were manageable with symptomatic treatments. Conclusion: For the treatment of advanced primary liver carcinoma, the combined therapy with oxaliplatin-based regimen and DC-CIK cells is effective, and its adverse effects are mild and well-tolerated, indicating that the therapy deserves further evaluation in more clinical trials with more patients.
Jiang Longwei
,
Huang Weiqian
,
Yao Lu
,
Zhang Yan
,
Ai Yueqin
,
Zheng Jie
,
Gao Yanrong
,
Zhang Chuang
,
Zhao Hua
,
Hu Jianhua
,
Jia Shaochang
,
Xiao Mei
2015, 22(6):765-772.
DOI: 10.3872/j.issn.1007-385X.2015.06.015
Abstract:
Objective:To evaluate the safety and clinical efficacy of the dendritic cells (DCs) and cytokine-induced killer (CIK) cells combination therapy in the treatment of moderate and advanced cervical cancer and analyze the prognostic factors. Methods: Thirty-nine patients with stage Ⅱb～Ⅳ cervical cancer who received DC-CIK cells treatment in the No. 81 hospital of PLA from Aug. 2011 to Dec. 2014 were enrolled into this retrospective analysis. Clinical response and changes of lymphocyte subsets and serum levels of tumor makers following the treatment were evaluated. Single and multiple-variable analyses were performed to identify the prognostic factors.Results: For the 39 patients received our DC-CIK cells immunotherapy, the overall response rate and the disease control rate were 20.5% and 66.7% respectively. Their 1-year overall survival rate was 61%, and both 2-year and 3-year survival rates were 46%. While there were no significant changes in lymphocyte subsets and SCCA levels in patients received the DC-CIK cells treatment, their CA125 levels were significantly decreased after after the therapy (51.79 vs 36.52, P<0.01). A single-factor analysis found that lymph node as well as distant metastasis can significantly affect the survival time of these patients. The multivariate Cox model indicated that age, lymph node metastasis, and the CA125 level before treatment were significantly associated with the risk of cancer-related death in the patients.Conclusion: Autologous DC-CIK cells immunotherapy is a safe and feasible treatment for patients with moderate and advanced cervical cancer and it may improve the long-term survival of these patients.
Objective:To evaluate the safety and clinical efficacy of the dendritic cells (DCs) and cytokine-induced killer (CIK) cells combination therapy in the treatment of moderate and advanced cervical cancer and analyze the prognostic factors. Methods: Thirty-nine patients with stage Ⅱb～Ⅳ cervical cancer who received DC-CIK cells treatment in the No. 81 hospital of PLA from Aug. 2011 to Dec. 2014 were enrolled into this retrospective analysis. Clinical response and changes of lymphocyte subsets and serum levels of tumor makers following the treatment were evaluated. Single and multiple-variable analyses were performed to identify the prognostic factors.Results: For the 39 patients received our DC-CIK cells immunotherapy, the overall response rate and the disease control rate were 20.5% and 66.7% respectively. Their 1-year overall survival rate was 61%, and both 2-year and 3-year survival rates were 46%. While there were no significant changes in lymphocyte subsets and SCCA levels in patients received the DC-CIK cells treatment, their CA125 levels were significantly decreased after after the therapy (51.79 vs 36.52, P<0.01). A single-factor analysis found that lymph node as well as distant metastasis can significantly affect the survival time of these patients. The multivariate Cox model indicated that age, lymph node metastasis, and the CA125 level before treatment were significantly associated with the risk of cancer-related death in the patients.Conclusion: Autologous DC-CIK cells immunotherapy is a safe and feasible treatment for patients with moderate and advanced cervical cancer and it may improve the long-term survival of these patients.
2015, 22(6):773-778.
DOI: 10.3872/j.issn.1007-385X.2015.06.016
Abstract:
Objective : To investigate the expression of KISS1 in peripheral blood and carcinoma tissues of patients with epithelial ovarian cancer (EOC) and its clinical implication. Methods: RT-PCR was used to assess KISS1 mRNA level in ovarian tissues and peripheral blood of 40 patients with EOC, 20 patients with borderline epithelial ovarian tumors, 20 patients with epithelial benign tumors, and 20 patients with hysteromyoma. The differences of KISS1 expression among the 4 types of tissues and the relationship between KISS1 mRNA level and clinicopathologic features of EOC patients were further analyzed. Results: The level of KISS1 mRNA in carcinoma tissues and peripheral blood of patients with EOC was significantly higher than that in benign tumors and normal control group (P<0.001). The expression of KISS1 in cancer tissues and peripheral blood of patients with EOC was associated with the tumor grade, clinical stage, metastasis, and the size of post-operation residual tumor (P<0.05). The level of KISS1 mRNA in primary tumors of EOC was markedly elevated compared with that in the corresponding metastatic lesions (P=0.001). Furthermore, KISS1 expression in ovarian tissues was positively correlated to that in the peripheral blood（r=0.669，P<0.001）. Conclusion: KISS1 gene may be involved in the development and progress of EOC. Peripheral blood KISS1 mRNA may serve as a biomarker for the differentiation status , stage, recurrence, and metastasis of ovarian cancer.
Objective : To investigate the expression of KISS1 in peripheral blood and carcinoma tissues of patients with epithelial ovarian cancer (EOC) and its clinical implication. Methods: RT-PCR was used to assess KISS1 mRNA level in ovarian tissues and peripheral blood of 40 patients with EOC, 20 patients with borderline epithelial ovarian tumors, 20 patients with epithelial benign tumors, and 20 patients with hysteromyoma. The differences of KISS1 expression among the 4 types of tissues and the relationship between KISS1 mRNA level and clinicopathologic features of EOC patients were further analyzed. Results: The level of KISS1 mRNA in carcinoma tissues and peripheral blood of patients with EOC was significantly higher than that in benign tumors and normal control group (P<0.001). The expression of KISS1 in cancer tissues and peripheral blood of patients with EOC was associated with the tumor grade, clinical stage, metastasis, and the size of post-operation residual tumor (P<0.05). The level of KISS1 mRNA in primary tumors of EOC was markedly elevated compared with that in the corresponding metastatic lesions (P=0.001). Furthermore, KISS1 expression in ovarian tissues was positively correlated to that in the peripheral blood（r=0.669，P<0.001）. Conclusion: KISS1 gene may be involved in the development and progress of EOC. Peripheral blood KISS1 mRNA may serve as a biomarker for the differentiation status , stage, recurrence, and metastasis of ovarian cancer.
2015, 22(6):779-784.
DOI: 10.3872/j.issn.1007-385X.2015.06.017
Abstract:
Objective:To examine the expression of phosphorylated mammalian target of rapamycin （p-mTOR） and hypoxia-inducible factor-1α(HIF-1α) in colorectal cancers and investigate its association with pathological features and prognosis.Methods：Surgical resected colorectal carcinoma and adjacent normal tissues were collected from patients with colorectal cancer enrolled into Jinan Military General Hospital from June, 2007 to December, 2013. Immunohistochemistry was performed to detect the expression of p-mTOR and HIF-1α in 63 cases of colorectal carcinoma tissues, 27 cases of metastatic lymph node tissues, and 12 cases of metastasized carcinoma tissues, with adjacent normal tissues as controls. χ2 test was used to analyze the relationship between p-mTOR and HIF-1α expression and clinical pathological features. Spearman correlation analysis was performed to analyze the association between p-mTOR and HIF-1α expression; Kaplan-Meier test was used for survival analysis related to p-mTOR and HIF-1α expression.Results: In colorectal carcinoma tissues, the positive rate of p-mTOR (50.80% vs 6.35%，P<0.01) and HIF-1α (65.08% vs 9.52%，P<0.01) was significantly higher than that of normal colorectal tissues. The expression of HIF-1α was correlated with distant metastasis，TNM stage and lymph node metastasis (P<0.05), and there was a positive correlation between p-mTOR and HIF-1α expression (r=0.345, P<0.01). Furthermore, the disease-free survival time and overall survival time of p-mTOR positive patients were significantly shorter than that of p-mTOR negative patients (P＜005). Conclusion: The expression of p-mTOR and HIF-1α are increased in colorectal cancer tissues, which are informative markers for clinical pathological staging and are useful prognosis factors.
Objective:To examine the expression of phosphorylated mammalian target of rapamycin （p-mTOR） and hypoxia-inducible factor-1α(HIF-1α) in colorectal cancers and investigate its association with pathological features and prognosis.Methods：Surgical resected colorectal carcinoma and adjacent normal tissues were collected from patients with colorectal cancer enrolled into Jinan Military General Hospital from June, 2007 to December, 2013. Immunohistochemistry was performed to detect the expression of p-mTOR and HIF-1α in 63 cases of colorectal carcinoma tissues, 27 cases of metastatic lymph node tissues, and 12 cases of metastasized carcinoma tissues, with adjacent normal tissues as controls. χ2 test was used to analyze the relationship between p-mTOR and HIF-1α expression and clinical pathological features. Spearman correlation analysis was performed to analyze the association between p-mTOR and HIF-1α expression; Kaplan-Meier test was used for survival analysis related to p-mTOR and HIF-1α expression.Results: In colorectal carcinoma tissues, the positive rate of p-mTOR (50.80% vs 6.35%，P<0.01) and HIF-1α (65.08% vs 9.52%，P<0.01) was significantly higher than that of normal colorectal tissues. The expression of HIF-1α was correlated with distant metastasis，TNM stage and lymph node metastasis (P<0.05), and there was a positive correlation between p-mTOR and HIF-1α expression (r=0.345, P<0.01). Furthermore, the disease-free survival time and overall survival time of p-mTOR positive patients were significantly shorter than that of p-mTOR negative patients (P＜005). Conclusion: The expression of p-mTOR and HIF-1α are increased in colorectal cancer tissues, which are informative markers for clinical pathological staging and are useful prognosis factors.
2015, 22(6):785-789.
DOI: 10.3872/j.issn.1007-385X.2015.06.018
Abstract:
Objective:To evaluate the clinical efficacy and safety of combined immunotherapy with dendritic cells (DCs) and cytokine-induced killer (CIK) cells in the treatment of relapsed multiple myeloma(MM). Methods: Bone marrow myeloma cells and peripheral blood mononuclear cells were collected from 22 cases of relapsed MM. The autologous DCs and CIK cells were amplified in vitro and used for reinfusion treatment. The clinical responses and immune parameters before and after the treatment were compared. Results: Among the 22 patients, 2 had CR in response to the combined therapy, 5 had nCR, 7 had PR, 3 had MR, and 5 had PD. Thus, the overall response rate (CR+nCR+PR) was 63.64% (14/22). After the treatment, the percentages of T cells subsets in peripheral blood did not change significantly (P>005), whereas the percentage of NK cells increased significantly (\[22.26±9.67\]% vs \[12.61±8.78\]%, P<001). Furthermore, the blood levels of LDH (\[166±41\]% vs \[112±21\]%) and β2-MG (\[5.11±2.33\]% vs \[265±1.61\]%) were significantly improved following the immunotherapy (P<0.01). Conclusion: The combined therapy with DC and CIK cells are safe and effective for relapsed MM. The treatment should be further tried in more patients with multiple myeloma and perhaps patients with B cell malignancies.
Objective:To evaluate the clinical efficacy and safety of combined immunotherapy with dendritic cells (DCs) and cytokine-induced killer (CIK) cells in the treatment of relapsed multiple myeloma(MM). Methods: Bone marrow myeloma cells and peripheral blood mononuclear cells were collected from 22 cases of relapsed MM. The autologous DCs and CIK cells were amplified in vitro and used for reinfusion treatment. The clinical responses and immune parameters before and after the treatment were compared. Results: Among the 22 patients, 2 had CR in response to the combined therapy, 5 had nCR, 7 had PR, 3 had MR, and 5 had PD. Thus, the overall response rate (CR+nCR+PR) was 63.64% (14/22). After the treatment, the percentages of T cells subsets in peripheral blood did not change significantly (P>005), whereas the percentage of NK cells increased significantly (\[22.26±9.67\]% vs \[12.61±8.78\]%, P<001). Furthermore, the blood levels of LDH (\[166±41\]% vs \[112±21\]%) and β2-MG (\[5.11±2.33\]% vs \[265±1.61\]%) were significantly improved following the immunotherapy (P<0.01). Conclusion: The combined therapy with DC and CIK cells are safe and effective for relapsed MM. The treatment should be further tried in more patients with multiple myeloma and perhaps patients with B cell malignancies.
Zhu Xuejun
,
Jiang Pengjun
,
Kong Xiangtu
,
Zhang Wenxi
,
Xia Wen
,
Fan Zhenfang
,
Yu Juhua
,
Sun Xuemei
,
Li Xiaohui
2015, 22(6):790-793.
DOI: 10.3872/j.issn.1007-385X.2015.06.019
Abstract:
近年来非霍奇金淋巴瘤（non-Hodgkin’s lymphoma，NHL）的发病率不断上升，化疗联合美罗华免疫治疗使NHL获得了较高的缓解率，但仍有部分患者因为原发耐药或者缓解后复发而无法获得治愈，因此研发新的淋巴瘤治疗方法对于提高疗效，延长患者生存具有重要的意义。近年来树突状细胞（dendritic cell, DC）疫苗免疫治疗各类肿瘤的基础与临床研究受到广泛的关注，通过给患者免疫接种携带肿瘤抗原的DC，可以诱导患者机体产生特异性的、持久的抗肿瘤免疫应答\[1-2\]；细胞因子诱导杀伤（cytokine induced killer，CIK）细胞输注可以调节和增强患者机体免疫功能\[3\]。因此本研究选择在淋巴瘤化疗完全缓解期，采用淋巴瘤抗原体外冲击的自体DC皮下免疫接种患者，其后静脉回输自体CIK细胞的方法免疫治疗B细胞淋巴瘤, 进行了初步的临床观察，现报道如下。
近年来非霍奇金淋巴瘤（non-Hodgkin’s lymphoma，NHL）的发病率不断上升，化疗联合美罗华免疫治疗使NHL获得了较高的缓解率，但仍有部分患者因为原发耐药或者缓解后复发而无法获得治愈，因此研发新的淋巴瘤治疗方法对于提高疗效，延长患者生存具有重要的意义。近年来树突状细胞（dendritic cell, DC）疫苗免疫治疗各类肿瘤的基础与临床研究受到广泛的关注，通过给患者免疫接种携带肿瘤抗原的DC，可以诱导患者机体产生特异性的、持久的抗肿瘤免疫应答\[1-2\]；细胞因子诱导杀伤（cytokine induced killer，CIK）细胞输注可以调节和增强患者机体免疫功能\[3\]。因此本研究选择在淋巴瘤化疗完全缓解期，采用淋巴瘤抗原体外冲击的自体DC皮下免疫接种患者，其后静脉回输自体CIK细胞的方法免疫治疗B细胞淋巴瘤, 进行了初步的临床观察，现报道如下。
2015, 22(6):794-798.
DOI: 10.3872/j.issn.1007-385X.2015.06.020
Abstract:
肿瘤基因突变的研究为转化医学研究提供了重要手段，对基因突变的深入探索改变了肿瘤的诊断、分类和预后评估方式，显著提高了肿瘤的临床治疗效果，同时也为个体化肿瘤免疫治疗的发展奠定基础并提供了新策略。本文对近年来这些领域的研究和肿瘤免疫治疗的临床应用进展予以介绍，并提出“突变组学(mutanome)”的概念，就基因突变组学与肿瘤的发生、与肿瘤疫苗治疗和过继免疫治疗的关系研究进展等方面作一综述。
肿瘤基因突变的研究为转化医学研究提供了重要手段，对基因突变的深入探索改变了肿瘤的诊断、分类和预后评估方式，显著提高了肿瘤的临床治疗效果，同时也为个体化肿瘤免疫治疗的发展奠定基础并提供了新策略。本文对近年来这些领域的研究和肿瘤免疫治疗的临床应用进展予以介绍，并提出“突变组学(mutanome)”的概念，就基因突变组学与肿瘤的发生、与肿瘤疫苗治疗和过继免疫治疗的关系研究进展等方面作一综述。
2015, 22(6):799-805.
DOI: 10.3872/j.issn.1007-385X.2015.06.021
Abstract:
胶质细胞系源性神经营养因子(glial cell line derived neurotrophic factor，GDNF)属于神经营养因子超家族，是TGF-β超家族的一个亚家族成员。目前已发现的GDNF家族配体(GDNF family ligand, GFL)有GDNF、neurturin (NRTN)、artemin (ARTN)、persephin (PSPN) 4个成员。GDNF家族受体为GFRα1-4，分别对应着GDNF家族配体GDNF、NRTN、ARTN及PSPN。通常GFRα通过糖基磷脂酰肌醇(glycosyl phosphatidyl inositol，GPI) 锚着到细胞膜上，经GDNF家族配体刺激后募集受体酪氨酸激酶RET作为共受体并形成GFL/GFRα/RET复合物，使RET酪氨酸激酶磷酸化，进一步活化下游的Ras/MAPK、PI3K、PLCγ信号通路进而调控细胞的功能。最初对GDNF家族成员的功能研究主要集中在神经系统，发现GDNF/GFRα信号对中枢神经系统有特异的营养作用和促轴突生长作用，在参与促进神经元存活及轴突损伤修复等方面有其他生长因子不可比拟的作用。但随着研究的进展，越来越多的资料表明GDNF/GFRα信号在肿瘤的发生发展中也占据一席之地，尤其在促进肿瘤侵袭转移方面具有重要作用。本文聚焦GDNF/GFRα1信号，重点阐述GDNF/ GFRα1信号在肿瘤进展和侵袭转移中的作用以及相关分子信号机制，以期为神经-内分泌-肿瘤相关性研究提供参考，为肿瘤的防治提供新的靶点和视角。
胶质细胞系源性神经营养因子(glial cell line derived neurotrophic factor，GDNF)属于神经营养因子超家族，是TGF-β超家族的一个亚家族成员。目前已发现的GDNF家族配体(GDNF family ligand, GFL)有GDNF、neurturin (NRTN)、artemin (ARTN)、persephin (PSPN) 4个成员。GDNF家族受体为GFRα1-4，分别对应着GDNF家族配体GDNF、NRTN、ARTN及PSPN。通常GFRα通过糖基磷脂酰肌醇(glycosyl phosphatidyl inositol，GPI) 锚着到细胞膜上，经GDNF家族配体刺激后募集受体酪氨酸激酶RET作为共受体并形成GFL/GFRα/RET复合物，使RET酪氨酸激酶磷酸化，进一步活化下游的Ras/MAPK、PI3K、PLCγ信号通路进而调控细胞的功能。最初对GDNF家族成员的功能研究主要集中在神经系统，发现GDNF/GFRα信号对中枢神经系统有特异的营养作用和促轴突生长作用，在参与促进神经元存活及轴突损伤修复等方面有其他生长因子不可比拟的作用。但随着研究的进展，越来越多的资料表明GDNF/GFRα信号在肿瘤的发生发展中也占据一席之地，尤其在促进肿瘤侵袭转移方面具有重要作用。本文聚焦GDNF/GFRα1信号，重点阐述GDNF/ GFRα1信号在肿瘤进展和侵袭转移中的作用以及相关分子信号机制，以期为神经-内分泌-肿瘤相关性研究提供参考，为肿瘤的防治提供新的靶点和视角。
2015, 22(6):806-810.
DOI: 10.3872/j.issn.1007-385X.2015.06.022
Abstract:
大肠癌是中国发病率排名第三的癌症。虽然近年来手术技术和术后化疗效果已经取得不小的进展，但大肠癌的预后仍不令人满意，主要原因是肿瘤的复发和转移。大肠癌干细胞Wnt信号通路在肿瘤复发和转移中起重要作用，并已成为抗癌药物研发的新靶点。大肠癌干细胞Wnt信号通路的失调导致细胞内β-catenin水平升高，继而激活一系列致癌相关基因。因此，如果针对Wnt信号通路的小分子靶向药物可阻止或逆转这些异常变化，将有助于治疗大肠癌。本文对以Wnt通路为靶标的小分子药物的研究现状进行了回顾，并展望其未来可能的发展方向。
大肠癌是中国发病率排名第三的癌症。虽然近年来手术技术和术后化疗效果已经取得不小的进展，但大肠癌的预后仍不令人满意，主要原因是肿瘤的复发和转移。大肠癌干细胞Wnt信号通路在肿瘤复发和转移中起重要作用，并已成为抗癌药物研发的新靶点。大肠癌干细胞Wnt信号通路的失调导致细胞内β-catenin水平升高，继而激活一系列致癌相关基因。因此，如果针对Wnt信号通路的小分子靶向药物可阻止或逆转这些异常变化，将有助于治疗大肠癌。本文对以Wnt通路为靶标的小分子药物的研究现状进行了回顾，并展望其未来可能的发展方向。
2015, 22(6):811-814.
DOI: 10.3872/j.issn.1007-385X.2015.06.023
Abstract:
microRNA(miRNA)是一类长度为20~24个核苷酸的非编码小RNA，它们可在转录后水平调节基因的表达。miRNA是许多细胞功能调控的关键因素，包括细胞的生长、增殖、分化及凋亡等。近来许多研究发现miR-133b与肿瘤的侵袭与转移密切相关，它可以通过抑制EGFR、fas、FSCN1及c-MET等多种基因的表达而发挥其抗肿瘤作用。本文主要对miR-133b在不同肿瘤中的异常表达及其可能的作用及其作用机制进行综述。
microRNA(miRNA)是一类长度为20~24个核苷酸的非编码小RNA，它们可在转录后水平调节基因的表达。miRNA是许多细胞功能调控的关键因素，包括细胞的生长、增殖、分化及凋亡等。近来许多研究发现miR-133b与肿瘤的侵袭与转移密切相关，它可以通过抑制EGFR、fas、FSCN1及c-MET等多种基因的表达而发挥其抗肿瘤作用。本文主要对miR-133b在不同肿瘤中的异常表达及其可能的作用及其作用机制进行综述。
2015, 22(6):815-818.
DOI: 10.3872/j.issn.1007-385X.2015.06.024
Abstract:
化疗药物耐药是影响肿瘤有效治疗的主要因素之一。5-氟尿嘧啶（5-fluorouracil，5-FU）作为一种基础化疗药物，其耐药机制成为研究热点。磷脂酰肌醇-3-激酶/蛋白激酶B（phosphatidylinositol-3-kinase/protein kinase B，PI3K/AKT）信号转导通路在促进细胞生长、运动、增殖、侵袭,抑制细胞凋亡，促进血管生成,抵抗化疗和放疗等方面起重要作用。近年来，关于PI3K/AKT信号通路与药物耐药性关系的研究越来越多，并被认为是化疗耐药治疗的新靶点。笔者查阅国内外近年来有关PI3K/AKT信号通路在结肠癌细胞5-FU耐药中作用机制的文献并做综述。
化疗药物耐药是影响肿瘤有效治疗的主要因素之一。5-氟尿嘧啶（5-fluorouracil，5-FU）作为一种基础化疗药物，其耐药机制成为研究热点。磷脂酰肌醇-3-激酶/蛋白激酶B（phosphatidylinositol-3-kinase/protein kinase B，PI3K/AKT）信号转导通路在促进细胞生长、运动、增殖、侵袭,抑制细胞凋亡，促进血管生成,抵抗化疗和放疗等方面起重要作用。近年来，关于PI3K/AKT信号通路与药物耐药性关系的研究越来越多，并被认为是化疗耐药治疗的新靶点。笔者查阅国内外近年来有关PI3K/AKT信号通路在结肠癌细胞5-FU耐药中作用机制的文献并做综述。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.