To investigate the roles of apoptosis and the Fas system in the process of liver cirrhosis converting into hepatocellular carcinoma , expression of Fas and Fas ligand in 49 LC and 36 HCC samples was detected by i...To investigate the roles of apoptosis and the Fas system in the process of liver cirrhosis converting into hepatocellular carcinoma , expression of Fas and Fas ligand in 49 LC and 36 HCC samples was detected by immunohistochemical method. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method. Serum soluble Fas levels in 28 cases of LC and 27 cases of HCC were measured by enzyme linked immunosorbent assay method. Compared with LC, apoptotic indices in HCC tissues were significantly reduced , expression of Fas was decreased , and that of FasL was increased . Serum sFas levels in HCC patients were significantly higher than those in normal controls. Down regulation of Fas expression, up regulation of FasL expression in hepatocytes and elevation of sFas level in serum might contribute to tumor escape from immune surveillance of the body. Apoptosis and the Fas system are significantly involved in the process of liver cirrhosis converting into hepatocellular carcinoma.展开更多
AIM: To study the in vitro and in vivo inhibitory effects of genistein on invasive potential of Bel 7402 hepatocellular carcinoma (HCC) cells and to explore the underlying mechanism.METHODS: Bel 7402 HCC cells were ex...AIM: To study the in vitro and in vivo inhibitory effects of genistein on invasive potential of Bel 7402 hepatocellular carcinoma (HCC) cells and to explore the underlying mechanism.METHODS: Bel 7402 HCC cells were exposed to genistein. The invasive activity of tumor cells was assayed in transwell cell culture chamber. p125FAK expression and cell cycle were evaluated by a functional assay. Cell apoptosis analysis was performed with TUNEL method. In addition, bilateral subrenal capsule xenograft transplantation of HCC was performed in 10 nude mice.Genistein was injected and the invasion of HCC into the renal parenchyma was observed. Microvessels with immunohistochemical staining were detected.RESULTS: Genistein significantly inhibited the growth of Bel 7402 cells, the inhibitory rate of tumor cells was 26 -42%. The invasive potential of Bel 7402 cells in vitro was significantly inhibited, the inhibitory rate was 11-28%. Genistein caused G2/M cell cycle arrest, S phase decreased significantly. The occurrence of apoptosis in genistein group increased significantly. The expression of p125FAK in 5 μg/mL genistein group (15.26±0.16%)and 10 μg/mL genistein group (12.89±0.36%) was significantly lower than that in the control group (19.75± 1.12%,P＜0.05). Tumor growth in genistein-treated nude mice was significantly retarded in comparison to control mice, the inhibitory rate of tumor growth was about 20%. Genistein also significantly inhibited the invasion of Bel 7402 cells into the renal parenchyma of nude mice with xenograft transplant. The positive unit value of microvessels in genistein-treated group (10.422±0.807)was significantly lower than that in control group (22.330 ± 5.696, P＜ 0.01).CONCLUSION: Genistein can effectively inhibit the invasive potential of Bel 7402 HCC cells by altering cell cycle, apoptosis and angiogenesis, inhibition of focal adhesion kinase may play a significant role in this process.展开更多
Background and Aims:DNA methylation and histone mod-ification are epigenetic modifications essential for normal function of mammalian cells.The processes are mediated by biochemical interactions between DNA methyltran...Background and Aims:DNA methylation and histone mod-ification are epigenetic modifications essential for normal function of mammalian cells.The processes are mediated by biochemical interactions between DNA methyltransferases(DNMTs)and histone deacetylases.Promoter hypermethyla-tion and deacetylation of tumor suppressor genes play major roles in cancer induction,through transcriptional silencing of these genes.DNA hypermethylation is carried out by a family of DNMTs including DNMT1,DNMT3a and DNMT3b.In hepatocellular carcinoma,a significant positive correlation bet-ween over-expression of these genes and cancer induction has been reported.The DNA demethylating agent genistein(GE)has been demonstrated to reduce different cancers.Pre-viously,we reported that GE can induce apoptosis and inhibit proliferation in hepatocellular carcinoma PLC/PRF5 and HepG2 cell lines.Besides,histone deacetylase inhibitors,such as trichostatin A(TSA),were successfully used to inhibit cancer cell growth.The present study was designed to assess the effect of GE in comparison with TSA on DNMT1,DNMT3a and DNMT3b gene expression,cell growth inhibition and apoptosis induction in the HepG2 cell line.Methods:Cells were seeded and treated with various doses of GE and TSA.The MTT assay,flow cytometry assay,and real-time RT-PCR were used to determine viability,apoptosis,and DNMT1,DNMT3a and DNMT3b gene expression respectively.Results:Both agents inhibited cell growth,induced apoptosis and re-activated DNMT1,DNMT3a and DNMT3b gene expression.Furthermore,TSA demonstrated a significantly greater apop-totic effect than the other agent,whereas GE improved gene expression more significantly than TSA.Conclusions:Our findings suggest that GE and TSA can significantly inhibit cell growth,induce apoptosis and restore DNMT1,DNMT3a and DNMT3b gene reactivation.展开更多
Objective：To investigate the effects of siRNA of ADAM17 gene and genistein on apoptosis and the inhibition of proliferation in HepG2 cells in an attempt to seek an effective therapy for hepatocellular carinoma. Meth...Objective：To investigate the effects of siRNA of ADAM17 gene and genistein on apoptosis and the inhibition of proliferation in HepG2 cells in an attempt to seek an effective therapy for hepatocellular carinoma. Methods：Cells were divided into control groups and experimental groups and siRNA was used to silence the ADAM17 gene, alone and in combination with genistein. Cells were harvested at several time periods and assessed for proliferation and apoptosis. Proliferation was assayed by MTT at 24, 48, 72 and 96 hours following treatment and apoptosis was assessed by flow cytometric analysis at 48 hours. Results：In siRNA groups, proliferation of cells was significantly inhibited compared to the control groups at 24, 48 and 72 hours（P 〈 0.05）, and apoptosis was significantly increased at 48 hours（P〈 0.01）; In genistein groups, proliferation was inhibited at 24, 48, 72 and 96 hours, and the apoptosis ratio was significantly increased at 48 hours（P〈 0.01）; while in the groups that received the combination of siRNA transfection and genistein treatment, there was a further significant decrease of proliferation and increase in apoptosis compared with either treatment alone. Conclusion：The ADAM17 gene could be an effective target, and genistein could be a useful agent, in the treatment of hepatocellular carcinoma, siRNA of ADAM17 gene and genistein both inhibited HepG2 cells proliferation and promoted apoptosis, and further, the combination of these treatments had a greater effect than either treatment alone.展开更多
基金This project was supported by Shanghai Science and Tech-nology Development Foundation(No.98XD140 2 2 )
文摘To investigate the roles of apoptosis and the Fas system in the process of liver cirrhosis converting into hepatocellular carcinoma , expression of Fas and Fas ligand in 49 LC and 36 HCC samples was detected by immunohistochemical method. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method. Serum soluble Fas levels in 28 cases of LC and 27 cases of HCC were measured by enzyme linked immunosorbent assay method. Compared with LC, apoptotic indices in HCC tissues were significantly reduced , expression of Fas was decreased , and that of FasL was increased . Serum sFas levels in HCC patients were significantly higher than those in normal controls. Down regulation of Fas expression, up regulation of FasL expression in hepatocytes and elevation of sFas level in serum might contribute to tumor escape from immune surveillance of the body. Apoptosis and the Fas system are significantly involved in the process of liver cirrhosis converting into hepatocellular carcinoma.
基金Supported by the Basic Research Key Project of the Science Foundation of Shanghai Municipal Commission of Science and Technology, No. 02JC14001
文摘AIM: To study the in vitro and in vivo inhibitory effects of genistein on invasive potential of Bel 7402 hepatocellular carcinoma (HCC) cells and to explore the underlying mechanism.METHODS: Bel 7402 HCC cells were exposed to genistein. The invasive activity of tumor cells was assayed in transwell cell culture chamber. p125FAK expression and cell cycle were evaluated by a functional assay. Cell apoptosis analysis was performed with TUNEL method. In addition, bilateral subrenal capsule xenograft transplantation of HCC was performed in 10 nude mice.Genistein was injected and the invasion of HCC into the renal parenchyma was observed. Microvessels with immunohistochemical staining were detected.RESULTS: Genistein significantly inhibited the growth of Bel 7402 cells, the inhibitory rate of tumor cells was 26 -42%. The invasive potential of Bel 7402 cells in vitro was significantly inhibited, the inhibitory rate was 11-28%. Genistein caused G2/M cell cycle arrest, S phase decreased significantly. The occurrence of apoptosis in genistein group increased significantly. The expression of p125FAK in 5 μg/mL genistein group (15.26±0.16%)and 10 μg/mL genistein group (12.89±0.36%) was significantly lower than that in the control group (19.75± 1.12%,P＜0.05). Tumor growth in genistein-treated nude mice was significantly retarded in comparison to control mice, the inhibitory rate of tumor growth was about 20%. Genistein also significantly inhibited the invasion of Bel 7402 cells into the renal parenchyma of nude mice with xenograft transplant. The positive unit value of microvessels in genistein-treated group (10.422±0.807)was significantly lower than that in control group (22.330 ± 5.696, P＜ 0.01).CONCLUSION: Genistein can effectively inhibit the invasive potential of Bel 7402 HCC cells by altering cell cycle, apoptosis and angiogenesis, inhibition of focal adhesion kinase may play a significant role in this process.
基金supported by adjutancy of research of Jahrom Medical University-Iran
文摘Background and Aims:DNA methylation and histone mod-ification are epigenetic modifications essential for normal function of mammalian cells.The processes are mediated by biochemical interactions between DNA methyltransferases(DNMTs)and histone deacetylases.Promoter hypermethyla-tion and deacetylation of tumor suppressor genes play major roles in cancer induction,through transcriptional silencing of these genes.DNA hypermethylation is carried out by a family of DNMTs including DNMT1,DNMT3a and DNMT3b.In hepatocellular carcinoma,a significant positive correlation bet-ween over-expression of these genes and cancer induction has been reported.The DNA demethylating agent genistein(GE)has been demonstrated to reduce different cancers.Pre-viously,we reported that GE can induce apoptosis and inhibit proliferation in hepatocellular carcinoma PLC/PRF5 and HepG2 cell lines.Besides,histone deacetylase inhibitors,such as trichostatin A(TSA),were successfully used to inhibit cancer cell growth.The present study was designed to assess the effect of GE in comparison with TSA on DNMT1,DNMT3a and DNMT3b gene expression,cell growth inhibition and apoptosis induction in the HepG2 cell line.Methods:Cells were seeded and treated with various doses of GE and TSA.The MTT assay,flow cytometry assay,and real-time RT-PCR were used to determine viability,apoptosis,and DNMT1,DNMT3a and DNMT3b gene expression respectively.Results:Both agents inhibited cell growth,induced apoptosis and re-activated DNMT1,DNMT3a and DNMT3b gene expression.Furthermore,TSA demonstrated a significantly greater apop-totic effect than the other agent,whereas GE improved gene expression more significantly than TSA.Conclusions:Our findings suggest that GE and TSA can significantly inhibit cell growth,induce apoptosis and restore DNMT1,DNMT3a and DNMT3b gene reactivation.
文摘Objective：To investigate the effects of siRNA of ADAM17 gene and genistein on apoptosis and the inhibition of proliferation in HepG2 cells in an attempt to seek an effective therapy for hepatocellular carinoma. Methods：Cells were divided into control groups and experimental groups and siRNA was used to silence the ADAM17 gene, alone and in combination with genistein. Cells were harvested at several time periods and assessed for proliferation and apoptosis. Proliferation was assayed by MTT at 24, 48, 72 and 96 hours following treatment and apoptosis was assessed by flow cytometric analysis at 48 hours. Results：In siRNA groups, proliferation of cells was significantly inhibited compared to the control groups at 24, 48 and 72 hours（P 〈 0.05）, and apoptosis was significantly increased at 48 hours（P〈 0.01）; In genistein groups, proliferation was inhibited at 24, 48, 72 and 96 hours, and the apoptosis ratio was significantly increased at 48 hours（P〈 0.01）; while in the groups that received the combination of siRNA transfection and genistein treatment, there was a further significant decrease of proliferation and increase in apoptosis compared with either treatment alone. Conclusion：The ADAM17 gene could be an effective target, and genistein could be a useful agent, in the treatment of hepatocellular carcinoma, siRNA of ADAM17 gene and genistein both inhibited HepG2 cells proliferation and promoted apoptosis, and further, the combination of these treatments had a greater effect than either treatment alone.
