Mobile website
Volume 18,Issue 3,2011 Table of Contents
2011, 18(3):239-243.
DOI: 10.3872/j.issn.1007-385X.2011.3.001
Abstract:
Cytokines, which can improve the host immunity and hamper the immune escape of tumor cells, have become an important strategy for tumor biotherapy. Interleukin (IL)-12 cytokine family is composed of heterodimeric cytokines, including IL-12, IL-23, IL-27 and IL-35; they are produced by antigen-presenting cells such as macrophages and dendritic cells and play critical roles in the differentiation of helper T (Th) cells. IL-12 induces IFN-γ production through activating signal transducer and activator of transcription (STAT) 4 signaling pathway and enhances cellular immune response; IL-27 augments cellular and humoral immune responses through activating STAT1 pathway; exogenous IL-23 can produce anti-tumor effects, while endogenous IL-23 induces the production of IL-17 by activating STAT3, promoting development and progress of tumors; and IL-35 may be a potential immunosuppressive cytokine. In summary, IL-12 cytokine family plays distinct roles in anti-tumor immune responses and has become a focus of study in the research and immune therapy of tumors.
Cytokines, which can improve the host immunity and hamper the immune escape of tumor cells, have become an important strategy for tumor biotherapy. Interleukin (IL)-12 cytokine family is composed of heterodimeric cytokines, including IL-12, IL-23, IL-27 and IL-35; they are produced by antigen-presenting cells such as macrophages and dendritic cells and play critical roles in the differentiation of helper T (Th) cells. IL-12 induces IFN-γ production through activating signal transducer and activator of transcription (STAT) 4 signaling pathway and enhances cellular immune response; IL-27 augments cellular and humoral immune responses through activating STAT1 pathway; exogenous IL-23 can produce anti-tumor effects, while endogenous IL-23 induces the production of IL-17 by activating STAT3, promoting development and progress of tumors; and IL-35 may be a potential immunosuppressive cytokine. In summary, IL-12 cytokine family plays distinct roles in anti-tumor immune responses and has become a focus of study in the research and immune therapy of tumors.
2011, 18(3):244-250.
DOI: 10.3872/j.issn.1007-385X.2011.3.002
Abstract:
Objective:To investigate the effects of melanoma differentiation associated gene-7 (mda-7, also named IL-24) and IL-24 delE5, an mda-7/IL-24 splice variant, on differentiation of acute myeloid leukemia (AML) cells. Methods: The AML cell lines U937 and HL-60 were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA); mda-7/IL-24 and IL-24 delE5 expressions were detected by real-time PCR and Western blotting assays. CD11b, CD14 and CD115 expressions on cell surface were examined by FACS. U937 and HL-60 cells were induced with TPA after siRNA interfering mda-7/IL-24 and IL-24 delE5 expressions, and then CD11b, CD14 and CD115 expressions were examined by FACS. U937 and HL-60 cell lines and primary leukemia cells from AML-M5 patients were transfected with mda-7/IL-24 or IL-24 delE5 plasmids, which were established in our previous study, in order to know whether ectopic overexpressions of mda-7/IL-24and IL-24 delE5 can induce the monocytic differentiation of leukemia cells. Results: The expressions of mda-7/IL-24 and its splice variant, IL-24 delE5, were significantly induced in U937 and HL-60 cells during TPA-mediated monocytic differentiation. Interfering mda-7/IL-24 and IL-24 delE5 expressions after siRNA treatment inhibited the monocytic differentiation ability of TPA on U937 and HL-60 cells. Ectopic overexpressions of mda-7/IL-24 and IL-24 delE5 induced differentiation of U937, HL-60, and primary AML leukemia cells into monocytes: CD11b, CD14 expressions on U937 and HL-60 cells and CD115 expression on U937 were significantly increased; CD11b, CD14 and CD115 expressions on primary leukemia cells from 3 AML patients were also increased with monocytic features. Conclusion: TPA can induce monocytic differentiation of leukemia cells by increasing mda-7/IL-24and its splice variant IL-24 delE5 expressions.
Objective:To investigate the effects of melanoma differentiation associated gene-7 (mda-7, also named IL-24) and IL-24 delE5, an mda-7/IL-24 splice variant, on differentiation of acute myeloid leukemia (AML) cells. Methods: The AML cell lines U937 and HL-60 were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA); mda-7/IL-24 and IL-24 delE5 expressions were detected by real-time PCR and Western blotting assays. CD11b, CD14 and CD115 expressions on cell surface were examined by FACS. U937 and HL-60 cells were induced with TPA after siRNA interfering mda-7/IL-24 and IL-24 delE5 expressions, and then CD11b, CD14 and CD115 expressions were examined by FACS. U937 and HL-60 cell lines and primary leukemia cells from AML-M5 patients were transfected with mda-7/IL-24 or IL-24 delE5 plasmids, which were established in our previous study, in order to know whether ectopic overexpressions of mda-7/IL-24and IL-24 delE5 can induce the monocytic differentiation of leukemia cells. Results: The expressions of mda-7/IL-24 and its splice variant, IL-24 delE5, were significantly induced in U937 and HL-60 cells during TPA-mediated monocytic differentiation. Interfering mda-7/IL-24 and IL-24 delE5 expressions after siRNA treatment inhibited the monocytic differentiation ability of TPA on U937 and HL-60 cells. Ectopic overexpressions of mda-7/IL-24 and IL-24 delE5 induced differentiation of U937, HL-60, and primary AML leukemia cells into monocytes: CD11b, CD14 expressions on U937 and HL-60 cells and CD115 expression on U937 were significantly increased; CD11b, CD14 and CD115 expressions on primary leukemia cells from 3 AML patients were also increased with monocytic features. Conclusion: TPA can induce monocytic differentiation of leukemia cells by increasing mda-7/IL-24and its splice variant IL-24 delE5 expressions.
2011, 18(3):251-257.
DOI: 10.3872/j.issn.1007-385X.2011.3.003
Abstract:
Objective:To observe the specific expression of tumstatin in hepatocarcinoma HepG2 cells induced by the human telomerase reverse transcriptase (hTERT) gene promoter and its antiangiogenic effect in vitro. Methods: phTERT-tumstatin, pCMV-tumstatin (positive control), phTERT-EGFP (negative control) plasmids were constructed and transfected into HepG2 and L-02 normal liver cells. The expression of EGFP was examined by fluorescence microscope. The expression of tumstatin protein in HepG2 cells was detected by Western blotting analysis; the proliferation of HepG2 cells after stably transfected with plasmids was measured by MTS assay; the effect of conditioned medium (containing tumstatin protein or not) on proliferation of human umbilical vascular endothelial cell (HUVEC) was detected by MTS assay. The effect of tumstatin protein on cellular tube structure formation of HUVEC was examined through counting the number of tube branches. Results: phTERT-tumstatin, pCMV-tumstatin, and phTERT-EGFP plasmids were successfully constructed. The specific expression of tumstatin was only observed in hepatocarcinoma HepG2, not in normal liver L-02 cells. phTERT-tumstatin and phTERT-EGFP transfection did not affect the proliferation of HepG2 cells; conditioned medium (CM) containing tumstatin protein (CM-T) inhibited the proliferation of HUVEC cells, with the inhibition rate being (56.49±0.33)%. The cellular tube structure formation of HUVEC cells on matrigel-coated plates supplemented with CM-T was significantly inhibited compared with conditioned medium CM-N and CM-NT (\[3.33±1.53\]% vs \[24.44±3.11\]%，\[23.94±2.92\]%, P<0.01). Conclusion: phTERT gene promoter can induce targeting expression of tumstatin in hepatocarcinoma cells and inhibit cellular tube structure formation of HUVEC cells.
Objective:To observe the specific expression of tumstatin in hepatocarcinoma HepG2 cells induced by the human telomerase reverse transcriptase (hTERT) gene promoter and its antiangiogenic effect in vitro. Methods: phTERT-tumstatin, pCMV-tumstatin (positive control), phTERT-EGFP (negative control) plasmids were constructed and transfected into HepG2 and L-02 normal liver cells. The expression of EGFP was examined by fluorescence microscope. The expression of tumstatin protein in HepG2 cells was detected by Western blotting analysis; the proliferation of HepG2 cells after stably transfected with plasmids was measured by MTS assay; the effect of conditioned medium (containing tumstatin protein or not) on proliferation of human umbilical vascular endothelial cell (HUVEC) was detected by MTS assay. The effect of tumstatin protein on cellular tube structure formation of HUVEC was examined through counting the number of tube branches. Results: phTERT-tumstatin, pCMV-tumstatin, and phTERT-EGFP plasmids were successfully constructed. The specific expression of tumstatin was only observed in hepatocarcinoma HepG2, not in normal liver L-02 cells. phTERT-tumstatin and phTERT-EGFP transfection did not affect the proliferation of HepG2 cells; conditioned medium (CM) containing tumstatin protein (CM-T) inhibited the proliferation of HUVEC cells, with the inhibition rate being (56.49±0.33)%. The cellular tube structure formation of HUVEC cells on matrigel-coated plates supplemented with CM-T was significantly inhibited compared with conditioned medium CM-N and CM-NT (\[3.33±1.53\]% vs \[24.44±3.11\]%，\[23.94±2.92\]%, P<0.01). Conclusion: phTERT gene promoter can induce targeting expression of tumstatin in hepatocarcinoma cells and inhibit cellular tube structure formation of HUVEC cells.
2011, 18(3):258-263.
DOI: 10.3872/j.issn.1007-385X.2011.3.004
Abstract:
Objective:To investigate the effect of tumor cells on lipid contents and protein secondary structures in dendritic cells (DCs) of different differentiation stages by Fourier transformed infrared spectrum (FT-IR), and to provide a clue for understanding the mechanism of tumor immune escape. Methods: CD14＋monocytes were isolated by immunomagnetic beads, and the immature DCs (imDCs) and mature DCs (mDCs) were induced by classic method. The imDCs and mDCs were co-cultured with human hepatoma BEL7402 cells, erythroleukemia K562 cells or human umbilical vein endothelial cells (HUVEC) for 48 h, untreated DCs served as control. The effects of tumor cells on lipid contents and protein secondary structures of DCs at different differentiation stages were investigated by FT-IR. Results: Compared with untreated DCs, the concentration of membrane phospholipid in DCs co-cultured with BEL7402 and K562 tumor cells was significantly decreased（2.718±0.296，3.124±0.361 vs 4.855±0.324，P<0.05; 2.964±0.136，3.522±0.173 vs 4.217±0.206，P<0.05）, the total lipid was increased （3.768±0.185，3.591±0.197 vs 2.487±0.212，P<0.05; 4.288±0.156，4.155±0.167 vs 3.233±0.206，P<0.05）, the protein α-helix was decreased （1.863±0.192，1.754±0.169 vs 2.364±0.188，P<0.05; 1.124±0.133，1.016±0.107 vs 1.392±0.113，P<0.05）， and the protein-sheet and β-turn were increased （3.397±0.225，3.433±0.236 vs 2.486±0.198，P<0.05; 2.646±0.209，2.591±0.216 vs1.558±0.159，P<0.05）. Moreover, compared with imDCs, mDCs were more susceptible to tumor-derived factors （4.366±0.284，4.322±0.266 vs 3.127±0.272，P<005; 2.675±0.221，2.627±0.235 vs 1.773±0.181，P<0.05）.Conclusion: Tumor cells can elicit the changes of lipid contents and protein secondary structures of imDCs and mDCs, which might be the structure basis for functional impairment caused by tumors.
Objective:To investigate the effect of tumor cells on lipid contents and protein secondary structures in dendritic cells (DCs) of different differentiation stages by Fourier transformed infrared spectrum (FT-IR), and to provide a clue for understanding the mechanism of tumor immune escape. Methods: CD14＋monocytes were isolated by immunomagnetic beads, and the immature DCs (imDCs) and mature DCs (mDCs) were induced by classic method. The imDCs and mDCs were co-cultured with human hepatoma BEL7402 cells, erythroleukemia K562 cells or human umbilical vein endothelial cells (HUVEC) for 48 h, untreated DCs served as control. The effects of tumor cells on lipid contents and protein secondary structures of DCs at different differentiation stages were investigated by FT-IR. Results: Compared with untreated DCs, the concentration of membrane phospholipid in DCs co-cultured with BEL7402 and K562 tumor cells was significantly decreased（2.718±0.296，3.124±0.361 vs 4.855±0.324，P<0.05; 2.964±0.136，3.522±0.173 vs 4.217±0.206，P<0.05）, the total lipid was increased （3.768±0.185，3.591±0.197 vs 2.487±0.212，P<0.05; 4.288±0.156，4.155±0.167 vs 3.233±0.206，P<0.05）, the protein α-helix was decreased （1.863±0.192，1.754±0.169 vs 2.364±0.188，P<0.05; 1.124±0.133，1.016±0.107 vs 1.392±0.113，P<0.05）， and the protein-sheet and β-turn were increased （3.397±0.225，3.433±0.236 vs 2.486±0.198，P<0.05; 2.646±0.209，2.591±0.216 vs1.558±0.159，P<0.05）. Moreover, compared with imDCs, mDCs were more susceptible to tumor-derived factors （4.366±0.284，4.322±0.266 vs 3.127±0.272，P<005; 2.675±0.221，2.627±0.235 vs 1.773±0.181，P<0.05）.Conclusion: Tumor cells can elicit the changes of lipid contents and protein secondary structures of imDCs and mDCs, which might be the structure basis for functional impairment caused by tumors.
2011, 18(3):264-269.
DOI: 10.3872/j.issn.1007-385X.2011.3.005
Abstract:
Objective:To investigate the apoptosis-inducing effect of an attenuated vesicular stomatitis virus in cervical cancer HeLa cells, and to explore the possible mechanism. Methods: HeLa cells were infected with VSV (MOI=1.0) and the cell proliferation was determined by MTT assay at 6, 12, 18, 24, and 30 h after infection. Morphological changes of apoptosis were observed by acridine orange (AO)/ethidium bromide (EB) staining. Annexin V/PI double-staining was performed to detect early apoptosis rate of HeLa cells and the sub-G1 apoptotic peak was examined by flow cytometry. The mitochondrial membrane potential of HeLa cells was measured by the JC-1 staining. The activities of caspase-9, -8 and -3 were measured by caspase assay kit. Results: After HeLa cells were exposed to attenuated VSV for 12 h and 24 h, the viabilities of cells were reduced to (78.4±1.9)% and (63.1±5.6)% (P<0.01); the early apoptosis rates were (16.88±248)% and (31.9±4.24)% (P<0.01); the Sub-G1 apoptotic peaks were (14.85±1.48)% and (21.05±228)% (P<0.01), respectively. Attenuated VSV significantly decreased mitochondrial membrane potential in HeLa cells with the increase of infection time (P<0.05). The activities of caspase-9 and caspase-3 of HeLa cells were significantly increased after VSV infection (all P<0.05). Conclusion: The attenuated VSV can inhibit the proliferation of HeLa cells and trigger apoptosis via caspase-9 and caspase-3-dependent pathway.
Objective:To investigate the apoptosis-inducing effect of an attenuated vesicular stomatitis virus in cervical cancer HeLa cells, and to explore the possible mechanism. Methods: HeLa cells were infected with VSV (MOI=1.0) and the cell proliferation was determined by MTT assay at 6, 12, 18, 24, and 30 h after infection. Morphological changes of apoptosis were observed by acridine orange (AO)/ethidium bromide (EB) staining. Annexin V/PI double-staining was performed to detect early apoptosis rate of HeLa cells and the sub-G1 apoptotic peak was examined by flow cytometry. The mitochondrial membrane potential of HeLa cells was measured by the JC-1 staining. The activities of caspase-9, -8 and -3 were measured by caspase assay kit. Results: After HeLa cells were exposed to attenuated VSV for 12 h and 24 h, the viabilities of cells were reduced to (78.4±1.9)% and (63.1±5.6)% (P<0.01); the early apoptosis rates were (16.88±248)% and (31.9±4.24)% (P<0.01); the Sub-G1 apoptotic peaks were (14.85±1.48)% and (21.05±228)% (P<0.01), respectively. Attenuated VSV significantly decreased mitochondrial membrane potential in HeLa cells with the increase of infection time (P<0.05). The activities of caspase-9 and caspase-3 of HeLa cells were significantly increased after VSV infection (all P<0.05). Conclusion: The attenuated VSV can inhibit the proliferation of HeLa cells and trigger apoptosis via caspase-9 and caspase-3-dependent pathway.
2011, 18(3):270-274.
DOI: 10.3872/j.issn.1007-385X.2011.3.006
Abstract:
Objective: To study the effect of triptolide (TP) on the proliferation of hepatocarcinoma SMMC-7721 cells and its effect on demethylation of P53 gene. Methods: The effect of TP on proliferation of SMMC-7721 cells was measured by MTT method, the effect of TP on P53 gene methylation in SMMC-7721 cells was analyzed by methylation specific PCR, the expressions of methyltransferase DNMT1, DNMT3a and DNMT3b mRNA in SMMC-7721 cells were measured by RT-PCR, and the protein expression of P53 in SMMC-7721 cells was detected by Western blotting assay. Results: TP inhibited the proliferation of SMMC-7721 cells in a dose-dependent manner (P<0.05), with the inhibitory rate being (73.5±3.02)% at 40 ng/ml TP, and IC50 of TP was 20 ng/ml. TP significantly inhibited DNMT1, DNMT3a, and DNMT3b mRNA expressions in SMMC-7721 cells (P<0.05, P<0.01), and dose-dependently reversed the hypermethylation of P53 gene and enhanced P53 protein expression in SMMC-7721 cells. Conclusion: TP can inhibit the proliferation of SMMC-7721 cells through the inhibiting methyltransferase expression, which augment the demethylation of P53 gene and results in the increased expression of P53 protein.
Objective: To study the effect of triptolide (TP) on the proliferation of hepatocarcinoma SMMC-7721 cells and its effect on demethylation of P53 gene. Methods: The effect of TP on proliferation of SMMC-7721 cells was measured by MTT method, the effect of TP on P53 gene methylation in SMMC-7721 cells was analyzed by methylation specific PCR, the expressions of methyltransferase DNMT1, DNMT3a and DNMT3b mRNA in SMMC-7721 cells were measured by RT-PCR, and the protein expression of P53 in SMMC-7721 cells was detected by Western blotting assay. Results: TP inhibited the proliferation of SMMC-7721 cells in a dose-dependent manner (P<0.05), with the inhibitory rate being (73.5±3.02)% at 40 ng/ml TP, and IC50 of TP was 20 ng/ml. TP significantly inhibited DNMT1, DNMT3a, and DNMT3b mRNA expressions in SMMC-7721 cells (P<0.05, P<0.01), and dose-dependently reversed the hypermethylation of P53 gene and enhanced P53 protein expression in SMMC-7721 cells. Conclusion: TP can inhibit the proliferation of SMMC-7721 cells through the inhibiting methyltransferase expression, which augment the demethylation of P53 gene and results in the increased expression of P53 protein.
2011, 18(3):275-279.
DOI: 10.3872/j.issn.1007-385X.2011.3.007
Abstract:
Objective:To study the effect of antisense human telomerase RNA (hTR) on adhesion and invasion of hepatocellular carcinoma (HCC) cell line BEL-7402 and its mechanism. Methods: Our previous research had successfully transfected sense or antisense hTR gene into BEL-7402 cells. This study they were divided into 3 groups: the sense hTR transfected group (BEL-7402-hTR-EcoRI), the antisense hTR transfected group (BEL-7402-hTR-BamHI) and the blank control group (BEL-7402). The adhesion and invasion of BEL-7402 cells after antisense hTR transfection were observed with adhesion and invasive assay, respectively. The expressions of integrin β1 (INTβ1) and epithelial cadherin (E-cad) were detected by immunocytochemistry, and galatin zymography was used to measure the activities of matrix metalloproteinase-2 (MMP-2) and MMP-9. Results: The adhesion and invasion abilities of antisense hTR transfected BEL-7402 cells were significantly weaker than those of the blank control and sense hTR transfected groups (0.204±0.029 vs 0.505±0037, 0.465±0.029; 34.80±3.19 vs 71.47±5.15, 69.87±3.11; all P<0.05). Meanwhile, the expression of INTβ1 was also significantly decreased (0.153 3±0.015 6 vs 0.385±0.008, 0.375±0.014 4, P<0.05), and E-cad was significantly increased (0.209±0.020 vs 0.124±0.018, 0.134±0.016; P<0.05) in the antisense hTR transfected group. The activities of MMP-2 and MMP-9 were significantly inhibited in the antisense hTR transfected group (2 05422±13852 vs 3 105.56±329.60, 2 923.22±269.08; 846.33±104.66 vs 1 538.89±122.85, 1 453.33±126.35; all P<0.05). Conclusion:Antisense hTR can inhibit the adhesion and invasion of hepatocellular carcinoma BEL-7402 cells, which may be related to the increase of E-cad expression, decrease of INTβ1 expression, and decrease of MMP-2 and MMP-9 activities.
Objective:To study the effect of antisense human telomerase RNA (hTR) on adhesion and invasion of hepatocellular carcinoma (HCC) cell line BEL-7402 and its mechanism. Methods: Our previous research had successfully transfected sense or antisense hTR gene into BEL-7402 cells. This study they were divided into 3 groups: the sense hTR transfected group (BEL-7402-hTR-EcoRI), the antisense hTR transfected group (BEL-7402-hTR-BamHI) and the blank control group (BEL-7402). The adhesion and invasion of BEL-7402 cells after antisense hTR transfection were observed with adhesion and invasive assay, respectively. The expressions of integrin β1 (INTβ1) and epithelial cadherin (E-cad) were detected by immunocytochemistry, and galatin zymography was used to measure the activities of matrix metalloproteinase-2 (MMP-2) and MMP-9. Results: The adhesion and invasion abilities of antisense hTR transfected BEL-7402 cells were significantly weaker than those of the blank control and sense hTR transfected groups (0.204±0.029 vs 0.505±0037, 0.465±0.029; 34.80±3.19 vs 71.47±5.15, 69.87±3.11; all P<0.05). Meanwhile, the expression of INTβ1 was also significantly decreased (0.153 3±0.015 6 vs 0.385±0.008, 0.375±0.014 4, P<0.05), and E-cad was significantly increased (0.209±0.020 vs 0.124±0.018, 0.134±0.016; P<0.05) in the antisense hTR transfected group. The activities of MMP-2 and MMP-9 were significantly inhibited in the antisense hTR transfected group (2 05422±13852 vs 3 105.56±329.60, 2 923.22±269.08; 846.33±104.66 vs 1 538.89±122.85, 1 453.33±126.35; all P<0.05). Conclusion:Antisense hTR can inhibit the adhesion and invasion of hepatocellular carcinoma BEL-7402 cells, which may be related to the increase of E-cad expression, decrease of INTβ1 expression, and decrease of MMP-2 and MMP-9 activities.
ZHANG Guo-jun
,
XU Meng
,
ZHAO Jian-fu
,
WANG Hong
,
XIANG Jun-jian
,
DENG Ning
,
ZENG Shi-bin
,
WANG Pan-pan
2011, 18(3):280-284.
DOI: 10.3872/j.issn.1007-385X.2011.3.008
Abstract:
Objective: To study the synergistic inhibitory effects of basic fibroblast growth factor (bFGF) monoclonal antibody (bFGF mAb ) and gimeracil and oteracil porassium (S-1) against proliferation of Lewis cells and the growth, metastasis, angiogenesis of the transplanted tumors. Methods: CCK-8 assay was used to assess the effects of bFGF mAb and S-1 on proliferation of Lewis cells. The spontaneous Lewis cell lung metastatic model was established, and thirty-two C57BL /6 mice were randomly divided into 4 groups: normal sodium (NS) group, bFGF mAb group, S-1 group, and bFGF mAb+S-1 group. Tumor volume was measured and tumor growth curve was drawn; tumors were weighed and the inhibitory rate of tumor growth was calculated; metastatic nodules on lung surface were counted; and the vascular endothelial cells were stained with CD31 to examine the microvessel density (MVD) of transplanted tumors. Results:Both bFGF mAb and S-1 inhibited Lewis cell proliferation in a dose-dependent manner (P<0.05). The inhibitory rate in bFGF mAb+S-1 group was significantly higher than those in the single drug treatment groups (P<0.05 or P<0.01). The inhibitory rates of transplanted tumors in bFGF mAb group, S-1 group, and bFGF mAb+S-1 groups were 37.8%, 47.7%, and 65.9%, respectively, with the combination group being significantly higher than the single treatment groups (P<0.05 or P<0.01). Moreover, the metastatic nodules and MVD in the combination group were significantly lower than those of single treatment groups (2.71±0.76 vs 6.57±0.98, 4.71±0.76; 21.6±2.9 vs 33.4±4.9, 41.9±63; P<0.05 or P<0.01). Conclusion: bFGF mAb and S-1 have synergistic inhibitory effects on Lewis transplanted tumors, which is related to the inhibition of proliferation and angiogenesis.
Objective: To study the synergistic inhibitory effects of basic fibroblast growth factor (bFGF) monoclonal antibody (bFGF mAb ) and gimeracil and oteracil porassium (S-1) against proliferation of Lewis cells and the growth, metastasis, angiogenesis of the transplanted tumors. Methods: CCK-8 assay was used to assess the effects of bFGF mAb and S-1 on proliferation of Lewis cells. The spontaneous Lewis cell lung metastatic model was established, and thirty-two C57BL /6 mice were randomly divided into 4 groups: normal sodium (NS) group, bFGF mAb group, S-1 group, and bFGF mAb+S-1 group. Tumor volume was measured and tumor growth curve was drawn; tumors were weighed and the inhibitory rate of tumor growth was calculated; metastatic nodules on lung surface were counted; and the vascular endothelial cells were stained with CD31 to examine the microvessel density (MVD) of transplanted tumors. Results:Both bFGF mAb and S-1 inhibited Lewis cell proliferation in a dose-dependent manner (P<0.05). The inhibitory rate in bFGF mAb+S-1 group was significantly higher than those in the single drug treatment groups (P<0.05 or P<0.01). The inhibitory rates of transplanted tumors in bFGF mAb group, S-1 group, and bFGF mAb+S-1 groups were 37.8%, 47.7%, and 65.9%, respectively, with the combination group being significantly higher than the single treatment groups (P<0.05 or P<0.01). Moreover, the metastatic nodules and MVD in the combination group were significantly lower than those of single treatment groups (2.71±0.76 vs 6.57±0.98, 4.71±0.76; 21.6±2.9 vs 33.4±4.9, 41.9±63; P<0.05 or P<0.01). Conclusion: bFGF mAb and S-1 have synergistic inhibitory effects on Lewis transplanted tumors, which is related to the inhibition of proliferation and angiogenesis.
2011, 18(3):285-289.
DOI: 10.3872/j.issn.1007-385X.2011.3.009
Abstract:
Objective:To transfect transcripted-carcinoembryonic antigen (CEA) mRNA into mature dendritic cells (DCs) in vitro, so as to observe the specific anti-tumor effect of DCs. Methods: Mature DCs was induced by GM-CSF, IL-4 TNF-α and then identified by FACS. pcDNA3.1-CEA plasmid was constructed and transcripted to CEA mRNA in vitro, and CEA mRNA was then tranfected into mature DCs by electroporation; CEA protein expression in DCs was examined by FACS, the proliferation of T cells induced by DCs was examined by MTT, the ability of DCs to induce specific anti-tumor responses of CTL was examined by LDH, and the level of IFN-γ in CTL supernatant was determined by ELISA. Results: CEA protein in CEA mRNA tranfected DCs was significantly increased compared with that in the control group (83.32% vs 3.34%, P<0.01). CEA mRNA tranfected DCs showed a stronger ability to induce the proliferation of T cells compared with control DCs when the ratio of effect cell to target cell (E∶T) being 1∶10 (\[19.11±1.89\]% vs \[1559±0.70\]%, P<0.05). CEA mRNA tranfected DCs induced specific anti-tumor responses of CTL, the cytotoxic rates being (5.42±0.87)%, (14.09±1.13)%, (27.16±0.72)%, and (32.49±0.84)% (P<0.01) when the E∶T were 5∶1, 10∶1, 20∶1 and 40∶1, and untransfected DCs and control target cells showed no cytotoxic effects. The level of IFN-γ in CTL supernatant induced by CEA mRNA tranfected DCs was significantly increased compared with that in the untransfected group (\[141.73±28.61\] pg/ml vs \[9.45±4.63\] pg/ml, P<0.01). Conclusion:Mature DCs transfected with CEA mRNA can induce specific anti-tumor responses, which provides a theoretical basis for CEA RNA-DCs vaccine.
Objective:To transfect transcripted-carcinoembryonic antigen (CEA) mRNA into mature dendritic cells (DCs) in vitro, so as to observe the specific anti-tumor effect of DCs. Methods: Mature DCs was induced by GM-CSF, IL-4 TNF-α and then identified by FACS. pcDNA3.1-CEA plasmid was constructed and transcripted to CEA mRNA in vitro, and CEA mRNA was then tranfected into mature DCs by electroporation; CEA protein expression in DCs was examined by FACS, the proliferation of T cells induced by DCs was examined by MTT, the ability of DCs to induce specific anti-tumor responses of CTL was examined by LDH, and the level of IFN-γ in CTL supernatant was determined by ELISA. Results: CEA protein in CEA mRNA tranfected DCs was significantly increased compared with that in the control group (83.32% vs 3.34%, P<0.01). CEA mRNA tranfected DCs showed a stronger ability to induce the proliferation of T cells compared with control DCs when the ratio of effect cell to target cell (E∶T) being 1∶10 (\[19.11±1.89\]% vs \[1559±0.70\]%, P<0.05). CEA mRNA tranfected DCs induced specific anti-tumor responses of CTL, the cytotoxic rates being (5.42±0.87)%, (14.09±1.13)%, (27.16±0.72)%, and (32.49±0.84)% (P<0.01) when the E∶T were 5∶1, 10∶1, 20∶1 and 40∶1, and untransfected DCs and control target cells showed no cytotoxic effects. The level of IFN-γ in CTL supernatant induced by CEA mRNA tranfected DCs was significantly increased compared with that in the untransfected group (\[141.73±28.61\] pg/ml vs \[9.45±4.63\] pg/ml, P<0.01). Conclusion:Mature DCs transfected with CEA mRNA can induce specific anti-tumor responses, which provides a theoretical basis for CEA RNA-DCs vaccine.
2011, 18(3):290-295.
DOI: 10.3872/j.issn.1007-385X.2011.3.010
Abstract:
Objective:To study the effect of siRNA silencing RhoC expression on apoptosis in human hepatocellular carcinoma BEL7402 cells and the related mechanism, so as to provide an experimental evidence for liver cancer gene therapy. Methods: RhoC-siRNA eukaryotic vector pU6mRFP RhoC-siRNA was constructed and transfected into BEL7402 cells, and the transfection efficiency was examined by confocal microscope. RT-PCR and Western blotting analysis were conducted to identify the effect of RhoC gene silencing; flow cytometry, agarose gel electrophoresis and Wright staining were applied to detect apoptosis of BEL7402 cells; and the expression levels of apoptosis related genes were determined by RT-PCR. Results: The pU6mRFP RhoC-siRNA recombinant plasmid was successfully constructed, and its transfection efficiency in BEL7402 cells was 70%. RT-PCR and Western blotting analysis results showed that the silencing rate of RhoC were 85% and 82%, respectively. The apoptosis rate of BEL7402 cells in pU6mRFP RhoC-siRNA transfected group was significantly higher than those in untransfected and pU6mRFP Scramble-siRNA transfected groups (\[21.00±2.23\]% vs \[6.47±164\]％,\[4.63±0.47\]%，P<0.01). Typical apoptotic DNA “ladder” appeared in pU6mRFP RhoC-siRNA transfected BEL7402 cells in gel electrophoresis analysis, and Wright staining showed a lot of apoptotic BEL7402 cells in pU6mRFP RhoC-siRNA group. Compared with untransfected and pU6mRFP Scramble-siRNA transfected BEL7402 cells, Bcl-2 gene expression in pU6mRFP RhoC-siRNA group was significantly decreased and Bax gene expression was significantly increased (0.28±0.15 vs 0.96±0.21, 1.03±0.24, P<0.05; 1.09±0.21 vs 0.26±0.10, 0.25±007, P<0.01). Conclusion: siRNA silencing RhoC gene expression can induce apoptosis in human hepatocellular carcinoma BEL7402 cells, which may be related to the down-regulated expression of Bcl-2 gene and up-regulated expression of Bax.
Objective:To study the effect of siRNA silencing RhoC expression on apoptosis in human hepatocellular carcinoma BEL7402 cells and the related mechanism, so as to provide an experimental evidence for liver cancer gene therapy. Methods: RhoC-siRNA eukaryotic vector pU6mRFP RhoC-siRNA was constructed and transfected into BEL7402 cells, and the transfection efficiency was examined by confocal microscope. RT-PCR and Western blotting analysis were conducted to identify the effect of RhoC gene silencing; flow cytometry, agarose gel electrophoresis and Wright staining were applied to detect apoptosis of BEL7402 cells; and the expression levels of apoptosis related genes were determined by RT-PCR. Results: The pU6mRFP RhoC-siRNA recombinant plasmid was successfully constructed, and its transfection efficiency in BEL7402 cells was 70%. RT-PCR and Western blotting analysis results showed that the silencing rate of RhoC were 85% and 82%, respectively. The apoptosis rate of BEL7402 cells in pU6mRFP RhoC-siRNA transfected group was significantly higher than those in untransfected and pU6mRFP Scramble-siRNA transfected groups (\[21.00±2.23\]% vs \[6.47±164\]％,\[4.63±0.47\]%，P<0.01). Typical apoptotic DNA “ladder” appeared in pU6mRFP RhoC-siRNA transfected BEL7402 cells in gel electrophoresis analysis, and Wright staining showed a lot of apoptotic BEL7402 cells in pU6mRFP RhoC-siRNA group. Compared with untransfected and pU6mRFP Scramble-siRNA transfected BEL7402 cells, Bcl-2 gene expression in pU6mRFP RhoC-siRNA group was significantly decreased and Bax gene expression was significantly increased (0.28±0.15 vs 0.96±0.21, 1.03±0.24, P<0.05; 1.09±0.21 vs 0.26±0.10, 0.25±007, P<0.01). Conclusion: siRNA silencing RhoC gene expression can induce apoptosis in human hepatocellular carcinoma BEL7402 cells, which may be related to the down-regulated expression of Bcl-2 gene and up-regulated expression of Bax.
2011, 18(3):296-300.
DOI: 10.3872/j.issn.1007-385X.2011.3.011
Abstract:
Objective:To study the effect of gemcitabine combined with TRAIL (TNF-related apoptosis inducing ligand) on proliferation and apoptosis of human lung cancer NCI-H460 cells. Methods: MTT assay was used to analyze the proliferation of NCI-H460 cells, and flow cytometry was used to examine the apoptosis of NCI-H460 cells. Results: Gemcitabine or TRAIL alone or in combination inhibited the proliferation of NCI-H460 cells in a time-(24, 48, 72 h) and concentration-dependent (0.001-10 μg/ml) manner. Gemcitabine combined with TRAIL had a stronger inhibitory effect than they were used alone, with the inhibitory ratios being 14.9%-77.5%, 23.4%-74.8%, and 22.3%-805% after 72 h in combination, gemcitabine, and TRAIL groups, and the IC50 being 0.0853, 0.0982, and 0.0435 μg/ml respectively. The inhibitory effect of gemcitabine combined with TRAIL on proliferation of NCI-H460 cells was related to the administration order and the ratio of gemcitabine and TRAIL; gemcitabine treatment for 24 h and then giving TRAIL had a stronger inhibitory effect than TRAIL treatment for 24 h and then giving gemcitabine; when the ratio of gemcitabine and TRAIL was 1:0.3, the combination showed the strongest inhibitory effect. The apoptosis rate of NCI-H460 cells in the combination treatment group was significantly higher than in the other two groups (49.04% vs 29.33%, 25.69%, P<0.01). Conclusion:Gemcitabine and TRAIL have synergistic effects in inhibiting proliferation and promoting apoptosis of lung cancer NCI-H460 cells.
Objective:To study the effect of gemcitabine combined with TRAIL (TNF-related apoptosis inducing ligand) on proliferation and apoptosis of human lung cancer NCI-H460 cells. Methods: MTT assay was used to analyze the proliferation of NCI-H460 cells, and flow cytometry was used to examine the apoptosis of NCI-H460 cells. Results: Gemcitabine or TRAIL alone or in combination inhibited the proliferation of NCI-H460 cells in a time-(24, 48, 72 h) and concentration-dependent (0.001-10 μg/ml) manner. Gemcitabine combined with TRAIL had a stronger inhibitory effect than they were used alone, with the inhibitory ratios being 14.9%-77.5%, 23.4%-74.8%, and 22.3%-805% after 72 h in combination, gemcitabine, and TRAIL groups, and the IC50 being 0.0853, 0.0982, and 0.0435 μg/ml respectively. The inhibitory effect of gemcitabine combined with TRAIL on proliferation of NCI-H460 cells was related to the administration order and the ratio of gemcitabine and TRAIL; gemcitabine treatment for 24 h and then giving TRAIL had a stronger inhibitory effect than TRAIL treatment for 24 h and then giving gemcitabine; when the ratio of gemcitabine and TRAIL was 1:0.3, the combination showed the strongest inhibitory effect. The apoptosis rate of NCI-H460 cells in the combination treatment group was significantly higher than in the other two groups (49.04% vs 29.33%, 25.69%, P<0.01). Conclusion:Gemcitabine and TRAIL have synergistic effects in inhibiting proliferation and promoting apoptosis of lung cancer NCI-H460 cells.
2011, 18(3):301-305.
DOI: 10.3872/j.issn.1007-385X.2011.3.012
Abstract:
Objective:To investigate changes of CD107a expression in γδT cells during cultivation and the relationship of CD107a expression with cytotoxicity of γδT cells. Methods: γδT cells were generated in vitro by stimulating PBMCs with IL-2 and isopentenyl pyrophosphate (IPP). Phenotype analysis of γδT cells was performed on the 7, 10 and 14 day by flow cytometry. Meanwhile, CD107a, perforin and granzyme B expressions were detected in γδT cells by flow cytometry. The cytotoxicity of γδT cells on pancreatic carcinoma SW-1990 cells was determined by CCK-8 kit. Spearman correlation analysis was performed by SPSS13.0 software. Results: γδTCR expression in γδT cells was (60.31±3.84)%, (66.45±4.25)% and (70.99±4.66)% on 7, 10 and 14 day, respectively. The expression of CD107a, perforin and granzyme B reached the peak on 7~10 d (7 d vs 0 d: \[80.66±4.42\]%, \[70.11±3.34\]%, \[94.26±4.25\]% vs \[69.02±5.04\]%, \[62.31±4.66\]%, \[53.62±3.69\]%, P<0.05), and then gradually decreased. The cytotoxicity rates of 7 day and 10 day γδT cells against SW-1990 cells were significantly higher than those of 14 day γδT cells (\[5886±5.12\]%, \[61.53±4.69\]% vs \[40.31±4.83\]%, P<0.05). CD107a expression in γδT cells was significantly correlated with perforin, granzyme B expressions and cytotoxicity on SW-1990 cells (P<0.01). Conclusion: The expression of CD107a on human peripheral γδT cells is positively correlated with its anti-tumor effect and may serve as a marker for the cytotoxic activity of γδT cells.
Objective:To investigate changes of CD107a expression in γδT cells during cultivation and the relationship of CD107a expression with cytotoxicity of γδT cells. Methods: γδT cells were generated in vitro by stimulating PBMCs with IL-2 and isopentenyl pyrophosphate (IPP). Phenotype analysis of γδT cells was performed on the 7, 10 and 14 day by flow cytometry. Meanwhile, CD107a, perforin and granzyme B expressions were detected in γδT cells by flow cytometry. The cytotoxicity of γδT cells on pancreatic carcinoma SW-1990 cells was determined by CCK-8 kit. Spearman correlation analysis was performed by SPSS13.0 software. Results: γδTCR expression in γδT cells was (60.31±3.84)%, (66.45±4.25)% and (70.99±4.66)% on 7, 10 and 14 day, respectively. The expression of CD107a, perforin and granzyme B reached the peak on 7~10 d (7 d vs 0 d: \[80.66±4.42\]%, \[70.11±3.34\]%, \[94.26±4.25\]% vs \[69.02±5.04\]%, \[62.31±4.66\]%, \[53.62±3.69\]%, P<0.05), and then gradually decreased. The cytotoxicity rates of 7 day and 10 day γδT cells against SW-1990 cells were significantly higher than those of 14 day γδT cells (\[5886±5.12\]%, \[61.53±4.69\]% vs \[40.31±4.83\]%, P<0.05). CD107a expression in γδT cells was significantly correlated with perforin, granzyme B expressions and cytotoxicity on SW-1990 cells (P<0.01). Conclusion: The expression of CD107a on human peripheral γδT cells is positively correlated with its anti-tumor effect and may serve as a marker for the cytotoxic activity of γδT cells.
2011, 18(3):306-310.
DOI: 10.3872/j.issn.1007-385X.2011.3.013
Abstract:
Objective:To screen differentially expressed microRNAs (miRNAs) in tumor and the corresponding paracancerous tissues of renal cell carcinoma (RCC) by massively parallel signature sequencing (MPSS), and to identify the expression of miR-660 in RCC. Methods: The expression of miRNAs in tumor and the corresponding paracancerous tissues of 10 RCC patients was examined by MPSS, and the differentially expressed miRNAs were screened. Expression of miR-660 in tumor tissues of 5 patients and the paracancerous tissues of 40 RCC patients were further examined by RT-PCR and qPCR, respectively. Results: The MPSS results showed that 283 miRNAs were up-regulated and 187 were down-regulated in RCC tissues, and the expression level of miR-660 in RCC tissues was only 19.5% of that in the paracancerous tissues. The MPSS results were primarily confirmed by RT-PCR, and qPCR results further showed that miR-660 expression in 34 of 40 RCC tissues was significantly lower than that in the paracancerous tissues, with the average level being only 11.7% of that in the paracancerous tissues. Conclusion: A total of 283 miRNAs are up-regulated and 187 down-regulated in RCC tissues; and miR-660 is lowly expressed in RCC, which may be a new target for the diagnosis and treatment of RCC.
Objective:To screen differentially expressed microRNAs (miRNAs) in tumor and the corresponding paracancerous tissues of renal cell carcinoma (RCC) by massively parallel signature sequencing (MPSS), and to identify the expression of miR-660 in RCC. Methods: The expression of miRNAs in tumor and the corresponding paracancerous tissues of 10 RCC patients was examined by MPSS, and the differentially expressed miRNAs were screened. Expression of miR-660 in tumor tissues of 5 patients and the paracancerous tissues of 40 RCC patients were further examined by RT-PCR and qPCR, respectively. Results: The MPSS results showed that 283 miRNAs were up-regulated and 187 were down-regulated in RCC tissues, and the expression level of miR-660 in RCC tissues was only 19.5% of that in the paracancerous tissues. The MPSS results were primarily confirmed by RT-PCR, and qPCR results further showed that miR-660 expression in 34 of 40 RCC tissues was significantly lower than that in the paracancerous tissues, with the average level being only 11.7% of that in the paracancerous tissues. Conclusion: A total of 283 miRNAs are up-regulated and 187 down-regulated in RCC tissues; and miR-660 is lowly expressed in RCC, which may be a new target for the diagnosis and treatment of RCC.
2011, 18(3):311-314.
DOI: 10.3872/j.issn.1007-385X.2011.3.014
Abstract:
Objective:To investigate the expression of CD14+DRlow/- myeloid-derived suppressor cells (MDSCs) in peripheral blood and tumor tissues of gastric carcinoma (GC) patients and its relationship with clinicopathological of GC. Methods: Forty-three stomach carcinoma patients (9 stageⅠ, 13 stageⅡ, 14 stageⅢ, 7stageⅣ) were selected from Third Affiliated Hospital of Anhui Medical University (Mar. 2009 to Oct. 2010), and 26 healthy volunteers were used as control. CD14+DRlow/-MDSCs expression in peripheral blood and gastric carcinoma tissues was detected by flow cytometry, and its relationship with clinicopathological of GC was analyzed. Results: CD14+DRlow/-MDSCs expression in GC tissues was significantly higher than those in peripheral blood of GC patients and healthy controls (\[2.87±1.93\]% vs \[237±1.7\]%, \[0.89±0.47\]%, P<0.05 and P<0.01), and CD14+DRlow/-MDSCs expression in peripheral blood of GC patients was also higher than that in healthy controls (P<0.01). CD14+DRlow/-MDSCs expression in GC tissues was positively correlated with their malignancy stage and significantly increased in advanced GC (stage Ⅰ∶Ⅱ∶Ⅲ∶Ⅳ=(1.15±0.78)%∶(1.71±0.92)%∶(2.25±1.24)%∶(4.85±2.37)%, P<0.05). Meanwhile, CD14+DRlow/-MDSCs expression in tumor infiltration tissues was significantly higher than that in tumor un-infiltration tissues (\[3.90±167)% vs \[2.62±1.53\]%, P<0.05). Conclusion: CD14+DRlow/-MDSCs are highly expressed in peripheral blood and GC tissues, and relates to the development and progression of GC.
Objective:To investigate the expression of CD14+DRlow/- myeloid-derived suppressor cells (MDSCs) in peripheral blood and tumor tissues of gastric carcinoma (GC) patients and its relationship with clinicopathological of GC. Methods: Forty-three stomach carcinoma patients (9 stageⅠ, 13 stageⅡ, 14 stageⅢ, 7stageⅣ) were selected from Third Affiliated Hospital of Anhui Medical University (Mar. 2009 to Oct. 2010), and 26 healthy volunteers were used as control. CD14+DRlow/-MDSCs expression in peripheral blood and gastric carcinoma tissues was detected by flow cytometry, and its relationship with clinicopathological of GC was analyzed. Results: CD14+DRlow/-MDSCs expression in GC tissues was significantly higher than those in peripheral blood of GC patients and healthy controls (\[2.87±1.93\]% vs \[237±1.7\]%, \[0.89±0.47\]%, P<0.05 and P<0.01), and CD14+DRlow/-MDSCs expression in peripheral blood of GC patients was also higher than that in healthy controls (P<0.01). CD14+DRlow/-MDSCs expression in GC tissues was positively correlated with their malignancy stage and significantly increased in advanced GC (stage Ⅰ∶Ⅱ∶Ⅲ∶Ⅳ=(1.15±0.78)%∶(1.71±0.92)%∶(2.25±1.24)%∶(4.85±2.37)%, P<0.05). Meanwhile, CD14+DRlow/-MDSCs expression in tumor infiltration tissues was significantly higher than that in tumor un-infiltration tissues (\[3.90±167)% vs \[2.62±1.53\]%, P<0.05). Conclusion: CD14+DRlow/-MDSCs are highly expressed in peripheral blood and GC tissues, and relates to the development and progression of GC.
2011, 18(3):315-319.
DOI: 10.3872/j.issn.1007-385X.2011.3.015
Abstract:
Objective: To study the ratio of circulating CD45low CD34+ KDR+ endothelial progenitor cells (EPCs) in peripheral blood of laryngeal squamous carcinoma patients, and to investigate its relationship with the development and progress of laryngeal squamous carcinoma. Methods: Blood samples from 20 laryngeal squamous carcinoma patients, who were diagnosed from Feb. 2010 to Feb. 2011, and 10 healthy volunteers were included in the present study. The ratio of CD45low CD34+ KDR+ EPCs in peripheral CD34+ haemopoietic stem cells or peripheral lymphocytes was detected by flow cytometry, the expressions of KDR and CD133 mRNA in PBMC were examined by real-time RT-PCR, and the levels of VEGF and NO in plasma were assayed by ELISA and Griess assay, respectively. Results: The ratios of CD45low CD34+ KDR+EPCs in both peripheral CD34+ haemopoietic stem cells and peripheral lymphocytes of laryngeal squamous carcinoma patients in stage Ⅱ-stage Ⅳ were significantly higher than those in healthy volunteers (\[55.4±11.4\]% vs \[21.0±5.8\]%，P<0.05; \[1.27±0.25\] vs \[0.25±0.09\], P<0.05). The expressions of KDR and CD133 mRNA and the level of plasma VEGF in laryngeal squamous carcinoma patients of stage Ⅱ-stage Ⅳ were significantly higher than those in healthy volunteers (P<0.05), but there was no significant difference in NO level. The ratio of CD45low CD34+ KDR+EPCs, the expression of KDR mRNA and CD133 mRNA, and the level of plasma VEGF in laryngeal squamous carcinoma patients of stageⅠwere similar to those in healthy volunteers. Conclusion: The ratio of circulating CD45low CD34+ KDR+ EPCs in peripheral blood and serum VEGF in advanced laryngeal squamous carcinoma patients are increased, which may be related to the development of tumors.
Objective: To study the ratio of circulating CD45low CD34+ KDR+ endothelial progenitor cells (EPCs) in peripheral blood of laryngeal squamous carcinoma patients, and to investigate its relationship with the development and progress of laryngeal squamous carcinoma. Methods: Blood samples from 20 laryngeal squamous carcinoma patients, who were diagnosed from Feb. 2010 to Feb. 2011, and 10 healthy volunteers were included in the present study. The ratio of CD45low CD34+ KDR+ EPCs in peripheral CD34+ haemopoietic stem cells or peripheral lymphocytes was detected by flow cytometry, the expressions of KDR and CD133 mRNA in PBMC were examined by real-time RT-PCR, and the levels of VEGF and NO in plasma were assayed by ELISA and Griess assay, respectively. Results: The ratios of CD45low CD34+ KDR+EPCs in both peripheral CD34+ haemopoietic stem cells and peripheral lymphocytes of laryngeal squamous carcinoma patients in stage Ⅱ-stage Ⅳ were significantly higher than those in healthy volunteers (\[55.4±11.4\]% vs \[21.0±5.8\]%，P<0.05; \[1.27±0.25\] vs \[0.25±0.09\], P<0.05). The expressions of KDR and CD133 mRNA and the level of plasma VEGF in laryngeal squamous carcinoma patients of stage Ⅱ-stage Ⅳ were significantly higher than those in healthy volunteers (P<0.05), but there was no significant difference in NO level. The ratio of CD45low CD34+ KDR+EPCs, the expression of KDR mRNA and CD133 mRNA, and the level of plasma VEGF in laryngeal squamous carcinoma patients of stageⅠwere similar to those in healthy volunteers. Conclusion: The ratio of circulating CD45low CD34+ KDR+ EPCs in peripheral blood and serum VEGF in advanced laryngeal squamous carcinoma patients are increased, which may be related to the development of tumors.
CHEN Ling
,
WANG Zong-hua
,
LI Xue-cheng
,
ZOU Li-quan
,
FAN Ya-chuan
,
LIU Zhi-peng
,
XIONG Xiao-feng
2011, 18(3):320-323.
DOI: 10.3872/j.issn.1007-385X.2011.3.016
Abstract:
Objective:To explore the expression of miR-29 in gastric cancer and its relationship with clinic pathological features and prognosis of gastric cancer. Methods: Totally 113 gastric cancer samples were obtained from patients who received operation during 2005 to 2010 in the No. 324 Hospital of PLA. The expression of miR-29 in gastric cancer and paracancerous tissues was detected by real-time PCR, and its relationship with clinic pathological features of gastric cancer was analyzed. Results: The expression of miR-29 in gastric cancer tissues was significantly lower than that in paracancerous tissues (2.62±1.17 vs 3.14±1.70, P=0.006). The rate of low miR-29 expression in patients with lymph node metastases was significantly higher than that without lymph node metastases (58.9% vs 32.5%, P=0.007). The median survival rate in low miR-29 expression cases was significantly lower than that in high miR-29 expression cases (33.0 vs 44.0 month, P=0.013). Conclusion: miR-29 is lowly expressed in gastric cancer tissues, and it might be used for prediction of prognosis and as for targeted treatment of gastric cancer.
Objective:To explore the expression of miR-29 in gastric cancer and its relationship with clinic pathological features and prognosis of gastric cancer. Methods: Totally 113 gastric cancer samples were obtained from patients who received operation during 2005 to 2010 in the No. 324 Hospital of PLA. The expression of miR-29 in gastric cancer and paracancerous tissues was detected by real-time PCR, and its relationship with clinic pathological features of gastric cancer was analyzed. Results: The expression of miR-29 in gastric cancer tissues was significantly lower than that in paracancerous tissues (2.62±1.17 vs 3.14±1.70, P=0.006). The rate of low miR-29 expression in patients with lymph node metastases was significantly higher than that without lymph node metastases (58.9% vs 32.5%, P=0.007). The median survival rate in low miR-29 expression cases was significantly lower than that in high miR-29 expression cases (33.0 vs 44.0 month, P=0.013). Conclusion: miR-29 is lowly expressed in gastric cancer tissues, and it might be used for prediction of prognosis and as for targeted treatment of gastric cancer.
2011, 18(3):324-326.
DOI: 10.3872/j.issn.1007-385X.2011.3.017
Abstract:
目的：探讨槲皮素和姜黄素对人肝癌HepG2细胞株增殖及凋亡的作用。方法：不同质量浓度的槲皮素（0、10、20、50、100 μmol/L）和姜黄素（0、2、5、10、20 μmol/L）分别作用HepG2细胞24、48、72 h，以As2O3（0、2、5、10、20 μmol/L）为阳性对照，MTT法检测HepG2细胞的增殖，流式细胞术检测HepG2细胞的凋亡。结果：槲皮素、姜黄素及As2O3作用后，HepG2细胞形态发生变化、生长速率变慢，HepG2增殖率随着时间和药物浓度的增加逐渐减低，72 h时3种药物高浓度组HepG2细胞增殖率分别为(31±5.8)%、(51±4.6)%、(54±5.8)%。槲皮素和姜黄素均有明显的促凋亡作用，并呈时间和浓度依赖性；高浓度(100 μmol/L)槲皮素作用最强，相同浓度下的姜黄素与As2O3作用相当。结论：槲皮素和姜黄素能明显抑制HepG2细胞增殖，促进其凋亡，并且这种作用强于As2O3。
目的：探讨槲皮素和姜黄素对人肝癌HepG2细胞株增殖及凋亡的作用。方法：不同质量浓度的槲皮素（0、10、20、50、100 μmol/L）和姜黄素（0、2、5、10、20 μmol/L）分别作用HepG2细胞24、48、72 h，以As2O3（0、2、5、10、20 μmol/L）为阳性对照，MTT法检测HepG2细胞的增殖，流式细胞术检测HepG2细胞的凋亡。结果：槲皮素、姜黄素及As2O3作用后，HepG2细胞形态发生变化、生长速率变慢，HepG2增殖率随着时间和药物浓度的增加逐渐减低，72 h时3种药物高浓度组HepG2细胞增殖率分别为(31±5.8)%、(51±4.6)%、(54±5.8)%。槲皮素和姜黄素均有明显的促凋亡作用，并呈时间和浓度依赖性；高浓度(100 μmol/L)槲皮素作用最强，相同浓度下的姜黄素与As2O3作用相当。结论：槲皮素和姜黄素能明显抑制HepG2细胞增殖，促进其凋亡，并且这种作用强于As2O3。
2011, 18(3):327-330.
DOI: 10.3872/j.issn.1007-385X.2011.3.018
Abstract:
目的：研究进展期肺腺癌患者血浆中EGFR基因突变与小分子酪氨酸激酶抑制剂（tyrosine kinase inhibitor，TKI）吉非替尼（gefitinib,又称易瑞沙Iressa）治疗进展期肺腺癌患者疗效间的关系。方法：选择上海交通大学医学院附属第三人民医院肿瘤科一线治疗失败后的44例进展期肺腺癌患者（男32例，女12例）和15位健康志愿者（男11例，女4例）。采集受试者血浆，利用改良酚-氯仿方法提取血浆DNA，利用突变富集型PCR（mutatant-enriched PCR，ME-PCR）扩增EGFR基因外显子19和21，然后进行基因测序。分析肺腺癌患者EGFR基因突变与与患者年龄、性别、吸烟史、体力状况（performance status，PS）评分、临床分期、是否接受放疗、一线化疗周期数间的关系，Kaplan-Meier法和Log-rank检验分析EGFR基因突变组与EGFR野生型组接受吉非替尼治疗的PFS和疗效差异。结果：44例肺腺癌患者血浆中有13例检出EGFR基因突变，其中外显子19缺失突变8例、外显子21点突变5例；健康对照组中未发现EGFR基因突变。肺腺癌患者EGFR基因突变与患者性别、吸烟史相关，与患者年龄、PS评分、分期、是否接受放疗及化疗周期数无关。EGFR基因突变的肺腺癌患者经吉非替尼治疗后中位无疾病进展时间(PFS)明显长于无EGFR突变患者（8个月vs 3.7个月，P=0.008）。结论：进展期肺腺癌患者血浆中EGFR基因突变检测可以作为分子靶向药物治疗的参考。
目的：研究进展期肺腺癌患者血浆中EGFR基因突变与小分子酪氨酸激酶抑制剂（tyrosine kinase inhibitor，TKI）吉非替尼（gefitinib,又称易瑞沙Iressa）治疗进展期肺腺癌患者疗效间的关系。方法：选择上海交通大学医学院附属第三人民医院肿瘤科一线治疗失败后的44例进展期肺腺癌患者（男32例，女12例）和15位健康志愿者（男11例，女4例）。采集受试者血浆，利用改良酚-氯仿方法提取血浆DNA，利用突变富集型PCR（mutatant-enriched PCR，ME-PCR）扩增EGFR基因外显子19和21，然后进行基因测序。分析肺腺癌患者EGFR基因突变与与患者年龄、性别、吸烟史、体力状况（performance status，PS）评分、临床分期、是否接受放疗、一线化疗周期数间的关系，Kaplan-Meier法和Log-rank检验分析EGFR基因突变组与EGFR野生型组接受吉非替尼治疗的PFS和疗效差异。结果：44例肺腺癌患者血浆中有13例检出EGFR基因突变，其中外显子19缺失突变8例、外显子21点突变5例；健康对照组中未发现EGFR基因突变。肺腺癌患者EGFR基因突变与患者性别、吸烟史相关，与患者年龄、PS评分、分期、是否接受放疗及化疗周期数无关。EGFR基因突变的肺腺癌患者经吉非替尼治疗后中位无疾病进展时间(PFS)明显长于无EGFR突变患者（8个月vs 3.7个月，P=0.008）。结论：进展期肺腺癌患者血浆中EGFR基因突变检测可以作为分子靶向药物治疗的参考。
2011, 18(3):331-336.
DOI: 10.3872/j.issn.1007-385X.2011.3.019
Abstract:
溶瘤病毒是一类具有复制能力的肿瘤杀伤性病毒，可通过多种机制发挥抗肿瘤作用。然而，机体的免疫监视作用使溶瘤病毒不能高效到达肿瘤部位而不足以治疗肿瘤。细胞载体作为溶瘤病毒的“特洛伊木马”可使溶瘤病毒逃避免疫监视，且两者联合具有更好的抗肿瘤疗效。根据细胞载体对肿瘤靶向程度的差异，细胞载体可分三大类：靶向肿瘤细胞的细胞载体，包括抗原特异性T细胞和细胞因子诱导的杀伤细胞；靶向肿瘤基质的细胞载体，包括间充质干细胞、内皮系细胞和肿瘤相关巨噬细胞；靶向组织或器官的细胞载体，包括外周血淋巴细胞、树突状细胞和肿瘤细胞等。本文主要介绍该三类细胞载体的研究及应用进展，讨论其存在的问题与应对对策，并展望未来发展方向。
溶瘤病毒是一类具有复制能力的肿瘤杀伤性病毒，可通过多种机制发挥抗肿瘤作用。然而，机体的免疫监视作用使溶瘤病毒不能高效到达肿瘤部位而不足以治疗肿瘤。细胞载体作为溶瘤病毒的“特洛伊木马”可使溶瘤病毒逃避免疫监视，且两者联合具有更好的抗肿瘤疗效。根据细胞载体对肿瘤靶向程度的差异，细胞载体可分三大类：靶向肿瘤细胞的细胞载体，包括抗原特异性T细胞和细胞因子诱导的杀伤细胞；靶向肿瘤基质的细胞载体，包括间充质干细胞、内皮系细胞和肿瘤相关巨噬细胞；靶向组织或器官的细胞载体，包括外周血淋巴细胞、树突状细胞和肿瘤细胞等。本文主要介绍该三类细胞载体的研究及应用进展，讨论其存在的问题与应对对策，并展望未来发展方向。
2011, 18(3):337-342.
DOI: 10.3872/j.issn.1007-385X.2011.3.020
Abstract:
间充质干细胞（mesenchymal stem cells，MSCs）是骨髓中具有自我更新和多向分化潜能的一种非造血干细胞，其免疫原性很低，在体外能抑制丝裂原或同种异基因抗原刺激的T细胞增殖。移植物抗宿主病（graft versus host disease，GVHD）是异基因造血干细胞移植（allogeneic hematopoietic stem cell transplantation，alloHSCT）后最主要的并发症，主要由供者T细胞所介导，是多种机制共同参与的复杂过程。在动物实验和临床试验中，MSCs具有较强的免疫调节作用，能显著抑制GVHD的发生，缓解其症状，提高受者的存活率；其机制为MSCs通过抑制T细胞的增殖，阻断DCs的分化和成熟，并通过增加CD4+CD25+调节性T细胞的比例来提高干细胞移植的成功率，使GVHD造成的病理损伤明显减轻，从而达到预防和治疗GVHD的目的。相关研究为MSCs有效应用于GVHD等临床免疫性疾病治疗提供了理论指导，并展现出极大的应用前景。
间充质干细胞（mesenchymal stem cells，MSCs）是骨髓中具有自我更新和多向分化潜能的一种非造血干细胞，其免疫原性很低，在体外能抑制丝裂原或同种异基因抗原刺激的T细胞增殖。移植物抗宿主病（graft versus host disease，GVHD）是异基因造血干细胞移植（allogeneic hematopoietic stem cell transplantation，alloHSCT）后最主要的并发症，主要由供者T细胞所介导，是多种机制共同参与的复杂过程。在动物实验和临床试验中，MSCs具有较强的免疫调节作用，能显著抑制GVHD的发生，缓解其症状，提高受者的存活率；其机制为MSCs通过抑制T细胞的增殖，阻断DCs的分化和成熟，并通过增加CD4+CD25+调节性T细胞的比例来提高干细胞移植的成功率，使GVHD造成的病理损伤明显减轻，从而达到预防和治疗GVHD的目的。相关研究为MSCs有效应用于GVHD等临床免疫性疾病治疗提供了理论指导，并展现出极大的应用前景。
2011, 18(3):343-346.
DOI: 10.3872/j.issn.1007-385X.2011.3.021
Abstract:
Cbl-b（casitas B-cell lineage lymphoma-b）为一种高度保守的RING家族E3泛素连接酶，是调节T细胞信号的关键因子，对维持外周T细胞免疫耐受至关重要。Cbl-b表达受转录因子、共刺激分子CD28、CTLA4及自身泛素化的调控。Cbl-b通过泛素化激活的受体、受体相关酪氨酸激酶和下游信号分子而参与淋巴细胞信号的负性调控，进而精密调节抗原受体信号和免疫反应。另外，Cbl-b在调控T细胞TGF-β信号转导通路中起着重要作用。Cbl-b的缺失能增强抗肿瘤免疫反应，打破自身免疫耐受。因此，深入研究Cbl-b在T细胞免疫耐受和肿瘤免疫治疗中的作用可为肿瘤免疫治疗提供新的靶点。
Cbl-b（casitas B-cell lineage lymphoma-b）为一种高度保守的RING家族E3泛素连接酶，是调节T细胞信号的关键因子，对维持外周T细胞免疫耐受至关重要。Cbl-b表达受转录因子、共刺激分子CD28、CTLA4及自身泛素化的调控。Cbl-b通过泛素化激活的受体、受体相关酪氨酸激酶和下游信号分子而参与淋巴细胞信号的负性调控，进而精密调节抗原受体信号和免疫反应。另外，Cbl-b在调控T细胞TGF-β信号转导通路中起着重要作用。Cbl-b的缺失能增强抗肿瘤免疫反应，打破自身免疫耐受。因此，深入研究Cbl-b在T细胞免疫耐受和肿瘤免疫治疗中的作用可为肿瘤免疫治疗提供新的靶点。
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
您是第位访问者
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.