目的:探讨细胞外的热休克蛋白7 0及H S P70肽复合物(h e a t s h o c k p r o t e i n 70/peptide complexes,HSP70/HSP70-PCs)对肝癌细胞HepG2缺氧诱导因子1(hypoxiainduciblefactor-1,HIF-1)、葡萄糖转运蛋白1(glucose transporter 1,...目的:探讨细胞外的热休克蛋白7 0及H S P70肽复合物(h e a t s h o c k p r o t e i n 70/peptide complexes,HSP70/HSP70-PCs)对肝癌细胞HepG2缺氧诱导因子1(hypoxiainduciblefactor-1,HIF-1)、葡萄糖转运蛋白1(glucose transporter 1,Glut1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响及可能机制.方法:将肝癌细胞分为3组:正常对照组、细胞外HSP70/HSP70-PCs诱导实验组(终浓度2g/mL)、细胞外HSP70/HSP70-PCs+siRNA转染组.应用Real-time RT-PCR与Western blot法检测HIF-1、Glut1和VEGF的表达变化.结果:细胞外HSP70/HSP70-PCs诱导实验组HIF-1、VEGF与Glut1蛋白表达显著升高,与对照组相比有显著差异(P<0.05),表明细胞外HSP70/HSP70-PCs促进肝癌细胞的HIF-1、Glut1和VEGF表达;应用siRNA阻断HIF-1后,细胞外HSP70/HSP70-PCs对Glut1和VEGF的上调表达作用消失,与对照组相比有明显差异(P<0.05).结论:细胞外HSP70/HSP70-PCs可以上调肝癌细胞HepG2中Glut1和VEGF表达,并且这一作用是通过HIF-1来实现的.展开更多
Apoptosis plays an important role in embryonic development, tissue remodeling, immune regulation and tumor regression. Two groups of molecules (Bcl-2 family and "Death factor" family) are involved in regulat...Apoptosis plays an important role in embryonic development, tissue remodeling, immune regulation and tumor regression. Two groups of molecules (Bcl-2 family and "Death factor" family) are involved in regulating apoptosis. In order to know about the effect of Bcl-2 on apoptosis induced by Fas, a typical member of "Death factor" family, the transfection experiments with expression vectors pcDNA3-fl and pcDNA3-bcl-2 were performed in BEL-7404 cells, a human hepatocellular carcinoma cell line which expresses endogenous Fas, but not FasL and Bcl-2. The data showed that the expression of FasL in pcDNA3-fl transfected hepatoma cells obviously induced the apoptosis of the cells. However, the overexpression of Bcl-2 in pcDNA3-bcl-2 transfected 7404/b-16 cells counteracted pcDNA3-fl transient transfection mediated apoptosis. Further study by cotransfection experiments indicated that Bid but not Bax (both were pro-apoptotic proteins of Bcl-2 family) blocked the inhibitory effect of Bcl-2 on Fas-mediated apoptosis. These results suggested that Fas-mediated apoptosis in human hepatoma cells is possibly regulated by Bcl-2 family proteins via mitochondria pathway.展开更多
AIM:To investigate the effect of the demethylating reagent 5-aza-2'-deoxycitidine(DAC) on telomerase activity in hepatocellular carcinoma(HCC) cell lines,SMMC-7721 and HepG2.METHODS:The related gene expression in ...AIM:To investigate the effect of the demethylating reagent 5-aza-2'-deoxycitidine(DAC) on telomerase activity in hepatocellular carcinoma(HCC) cell lines,SMMC-7721 and HepG2.METHODS:The related gene expression in cell lines was examined by real-time reverse transcription-polymerase chain reaction and Western blotting analysis.The telomerase activity was examined by telomeric repeat amplification protocol-enzyme-linked immunosorbent assay and DNA methylation was determined by methylation-specific polymerase chain reaction.RESULTS:The telomerase activity was significantly reduced in both cell lines treated with DAC,accompanied by downregulation of telomerase reverse transcriptase(hTERT).We also observed the effect of DAC on the methylation status of hTERT promoter and the expression of regulatory genes,such as c-myc,p15,p16,p21,E2F1,and WT1.The methylation status of hTERT promoter could be reversed in SMMC-7721 by DAC,but not in HepG2 cells.However,p16 expression could be reactivated by demethylation of its promoter,and c-Myc expression was repressed in both cell lines.Moreover,DAC could enhance the sensitivity to the chemotherapeutic agents,such as cisplatin,by induction of apoptosis of HCC cells.CONCLUSION:The DAC exerts its anti-tumor effects in HCC cells by inhibiting the telomerase activity.展开更多
AIM:To compare the clinical outcome and pathologic features of non-alcoholic steatohepatitis(NASH) patients with hepatocellular carcinoma(HCC) and hepatitic C virus(HCV) patients with HCC(another group in which HCC is...AIM:To compare the clinical outcome and pathologic features of non-alcoholic steatohepatitis(NASH) patients with hepatocellular carcinoma(HCC) and hepatitic C virus(HCV) patients with HCC(another group in which HCC is commonly seen) undergoing liver transplantation.METHODS:Patients transplanted for HCV and NASH at our institution from January 2000 to April 2011 were analyzed.All explanted liver histology and pre-transplant liver biopsies were examined by two specialist liver histopathologists.Patient demographics,disease free survival,explant liver characteristics and HCC features(tumour number,cumulative tumour size,vascular invasion and differentiation) were compared between HCV and NASH liver transplant recipients.RESULTS:A total of 102 patients with NASH and 283 patients with HCV were transplanted.The incidence of HCC in NASH transplant recipients was 16.7%(17/102).The incidence of HCC in HCV transplant recipients was 22.6%(64/283).Patients with NASH-HCC were statistically older than HCV-HCC patients(P < 0.001).A significantly higher proportion of HCV-HCC patients had vascular invasion(23.4% vs 6.4%,P = 0.002) and poorly differentiated HCC(4.7% vs 0%,P < 0.001) compared to the NASH-HCC group.A trend of poorer recurrence free survival at 5 years was seen in HCV-HCC patients compared to NASH-HCC who underwent a Liver transplantation(P = 0.11).CONCLUSION:Patients transplanted for NASH-HCC appear to have less aggressive tumour features compared to those with HCV-HCC,which likely in part accounts for their improved recurrence free survival.展开更多
AIM To investigate the clinicopathological significance of progesterone receptor membrane component 1(PGRMC1) and PGRMC2 in hepatocellular carcinoma(HCC). METHODS We performed immunohistochemical staining to evaluate ...AIM To investigate the clinicopathological significance of progesterone receptor membrane component 1(PGRMC1) and PGRMC2 in hepatocellular carcinoma(HCC). METHODS We performed immunohistochemical staining to evaluate the estrogen receptor(ER), progesterone receptor(PR), PGRMC1, and PGRMC2 in a clinical cohort consisting of 89 paired HCC and non-tumor liver samples. We also analyzed HCC data(n = 373) from The Cancer Genome Atlas(TCGA). We correlated the expression status of PGRMC1 and PGRMC2 with clinicopathological indicators and the clinical outcomes of the HCC patients. We knocked down or overexpressed PGRMC1 in HCC cell lines to evaluate its biological significance in HCC cell proliferation, differentiation, migration, and invasion. RESULTS We found that few HCC cases expressed ER(5.6%) and PR(4.5%). In contrast, most HCC cases expressed PGRMC1(89.9%) and PGRMC2(100%). PGRMC1 and PGRMC2 exhibited significantly lower expression in tumor tissue than in non-tumor tissue(P < 0.001). Lower PGRMC1 expression in HCC was significantly associated with higher serum alpha-fetoprotein expression(P = 0.004), poorer tumor differentiation(P = 0.045) and liver capsule penetration(P = 0.038). Low PGRMC1 expression was an independent predictor for worse disease-free survival(P = 0.002, HR = 2.384,CI: 1.377-4.128) in our cases, as well as in the TCGA cohort(P < 0.001, HR = 2.857, CI: 1.781-4.584). The expression of PGRMC2 did not relate to patient outcome. PGRMC1 knockdown promoted a poorly differentiated phenotype and proliferation of HCC cells in vitro, while PGRMC1 overexpression caused the opposite effects.CONCLUSION PGRMC1 is a non-classical hormonal receptor that negatively regulates hepatocarcinogenesis. PGRMC1 down-regulation is associated with progression of HCC and is a poor prognostic indicator.展开更多
Five types of superparamagnetic iron oxide （SPIO）,i.e. Ferumoxides （Feridex？ Ⅳ, Berlex Laboratories）,Fe r u c a r b o t ra n （ Re s ov i s t？, B aye r H e a l t h c a re ） ,Ferumoxtran-10 （AMI-227 or Code-72...Five types of superparamagnetic iron oxide （SPIO）,i.e. Ferumoxides （Feridex？ Ⅳ, Berlex Laboratories）,Fe r u c a r b o t ra n （ Re s ov i s t？, B aye r H e a l t h c a re ） ,Ferumoxtran-10 （AMI-227 or Code-7227, Combidex？, AMAG Pharma; Sinerem？, Guerbet）, NC100150（Clariscan？, Nycomed,） and （VSOP C184, Ferropharm）have been designed and clinically tested as magneticresonance contrast agents. However, until nowResovist？ is current available in only a few countries.The other four agents have been stopped for furtherdevelopment or withdrawn from the market. AnotherSPIO agent Ferumoxytol （Feraheme） is approved forthe treatment of iron deficiency in adult chronic kidneydisease patients. Ferumoxytol is comprised of ironoxide particles surrounded by a carbohydrate coat, andit is being explored as a potential imaging approach forevaluating lymph nodes and certain liver tumors.展开更多
文摘目的:探讨细胞外的热休克蛋白7 0及H S P70肽复合物(h e a t s h o c k p r o t e i n 70/peptide complexes,HSP70/HSP70-PCs)对肝癌细胞HepG2缺氧诱导因子1(hypoxiainduciblefactor-1,HIF-1)、葡萄糖转运蛋白1(glucose transporter 1,Glut1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响及可能机制.方法:将肝癌细胞分为3组:正常对照组、细胞外HSP70/HSP70-PCs诱导实验组(终浓度2g/mL)、细胞外HSP70/HSP70-PCs+siRNA转染组.应用Real-time RT-PCR与Western blot法检测HIF-1、Glut1和VEGF的表达变化.结果:细胞外HSP70/HSP70-PCs诱导实验组HIF-1、VEGF与Glut1蛋白表达显著升高,与对照组相比有显著差异(P<0.05),表明细胞外HSP70/HSP70-PCs促进肝癌细胞的HIF-1、Glut1和VEGF表达;应用siRNA阻断HIF-1后,细胞外HSP70/HSP70-PCs对Glut1和VEGF的上调表达作用消失,与对照组相比有明显差异(P<0.05).结论:细胞外HSP70/HSP70-PCs可以上调肝癌细胞HepG2中Glut1和VEGF表达,并且这一作用是通过HIF-1来实现的.
基金Major State Basic Reaearch (973) Program of China.
文摘Apoptosis plays an important role in embryonic development, tissue remodeling, immune regulation and tumor regression. Two groups of molecules (Bcl-2 family and "Death factor" family) are involved in regulating apoptosis. In order to know about the effect of Bcl-2 on apoptosis induced by Fas, a typical member of "Death factor" family, the transfection experiments with expression vectors pcDNA3-fl and pcDNA3-bcl-2 were performed in BEL-7404 cells, a human hepatocellular carcinoma cell line which expresses endogenous Fas, but not FasL and Bcl-2. The data showed that the expression of FasL in pcDNA3-fl transfected hepatoma cells obviously induced the apoptosis of the cells. However, the overexpression of Bcl-2 in pcDNA3-bcl-2 transfected 7404/b-16 cells counteracted pcDNA3-fl transient transfection mediated apoptosis. Further study by cotransfection experiments indicated that Bid but not Bax (both were pro-apoptotic proteins of Bcl-2 family) blocked the inhibitory effect of Bcl-2 on Fas-mediated apoptosis. These results suggested that Fas-mediated apoptosis in human hepatoma cells is possibly regulated by Bcl-2 family proteins via mitochondria pathway.
基金Supported by The National Natural Science Foundation of China,No.30901722,30973433,81000970,81030041,31171321 and 81101622
文摘AIM:To investigate the effect of the demethylating reagent 5-aza-2'-deoxycitidine(DAC) on telomerase activity in hepatocellular carcinoma(HCC) cell lines,SMMC-7721 and HepG2.METHODS:The related gene expression in cell lines was examined by real-time reverse transcription-polymerase chain reaction and Western blotting analysis.The telomerase activity was examined by telomeric repeat amplification protocol-enzyme-linked immunosorbent assay and DNA methylation was determined by methylation-specific polymerase chain reaction.RESULTS:The telomerase activity was significantly reduced in both cell lines treated with DAC,accompanied by downregulation of telomerase reverse transcriptase(hTERT).We also observed the effect of DAC on the methylation status of hTERT promoter and the expression of regulatory genes,such as c-myc,p15,p16,p21,E2F1,and WT1.The methylation status of hTERT promoter could be reversed in SMMC-7721 by DAC,but not in HepG2 cells.However,p16 expression could be reactivated by demethylation of its promoter,and c-Myc expression was repressed in both cell lines.Moreover,DAC could enhance the sensitivity to the chemotherapeutic agents,such as cisplatin,by induction of apoptosis of HCC cells.CONCLUSION:The DAC exerts its anti-tumor effects in HCC cells by inhibiting the telomerase activity.
文摘AIM:To compare the clinical outcome and pathologic features of non-alcoholic steatohepatitis(NASH) patients with hepatocellular carcinoma(HCC) and hepatitic C virus(HCV) patients with HCC(another group in which HCC is commonly seen) undergoing liver transplantation.METHODS:Patients transplanted for HCV and NASH at our institution from January 2000 to April 2011 were analyzed.All explanted liver histology and pre-transplant liver biopsies were examined by two specialist liver histopathologists.Patient demographics,disease free survival,explant liver characteristics and HCC features(tumour number,cumulative tumour size,vascular invasion and differentiation) were compared between HCV and NASH liver transplant recipients.RESULTS:A total of 102 patients with NASH and 283 patients with HCV were transplanted.The incidence of HCC in NASH transplant recipients was 16.7%(17/102).The incidence of HCC in HCV transplant recipients was 22.6%(64/283).Patients with NASH-HCC were statistically older than HCV-HCC patients(P < 0.001).A significantly higher proportion of HCV-HCC patients had vascular invasion(23.4% vs 6.4%,P = 0.002) and poorly differentiated HCC(4.7% vs 0%,P < 0.001) compared to the NASH-HCC group.A trend of poorer recurrence free survival at 5 years was seen in HCV-HCC patients compared to NASH-HCC who underwent a Liver transplantation(P = 0.11).CONCLUSION:Patients transplanted for NASH-HCC appear to have less aggressive tumour features compared to those with HCV-HCC,which likely in part accounts for their improved recurrence free survival.
基金Supported by the Ministry of Science and Technology,No.NSC102-2320-B-006-011.,No.MOST103-2320-B-006-021-MY2,and No.MOST105-2320-B-006-033 to Tsai HWNational Cheng Kung University Hospital,Taiwan,No.NCKUH-10406002 and No.NCKUH-10509001 to Tsai HW
文摘AIM To investigate the clinicopathological significance of progesterone receptor membrane component 1(PGRMC1) and PGRMC2 in hepatocellular carcinoma(HCC). METHODS We performed immunohistochemical staining to evaluate the estrogen receptor(ER), progesterone receptor(PR), PGRMC1, and PGRMC2 in a clinical cohort consisting of 89 paired HCC and non-tumor liver samples. We also analyzed HCC data(n = 373) from The Cancer Genome Atlas(TCGA). We correlated the expression status of PGRMC1 and PGRMC2 with clinicopathological indicators and the clinical outcomes of the HCC patients. We knocked down or overexpressed PGRMC1 in HCC cell lines to evaluate its biological significance in HCC cell proliferation, differentiation, migration, and invasion. RESULTS We found that few HCC cases expressed ER(5.6%) and PR(4.5%). In contrast, most HCC cases expressed PGRMC1(89.9%) and PGRMC2(100%). PGRMC1 and PGRMC2 exhibited significantly lower expression in tumor tissue than in non-tumor tissue(P < 0.001). Lower PGRMC1 expression in HCC was significantly associated with higher serum alpha-fetoprotein expression(P = 0.004), poorer tumor differentiation(P = 0.045) and liver capsule penetration(P = 0.038). Low PGRMC1 expression was an independent predictor for worse disease-free survival(P = 0.002, HR = 2.384,CI: 1.377-4.128) in our cases, as well as in the TCGA cohort(P < 0.001, HR = 2.857, CI: 1.781-4.584). The expression of PGRMC2 did not relate to patient outcome. PGRMC1 knockdown promoted a poorly differentiated phenotype and proliferation of HCC cells in vitro, while PGRMC1 overexpression caused the opposite effects.CONCLUSION PGRMC1 is a non-classical hormonal receptor that negatively regulates hepatocarcinogenesis. PGRMC1 down-regulation is associated with progression of HCC and is a poor prognostic indicator.
文摘Five types of superparamagnetic iron oxide （SPIO）,i.e. Ferumoxides （Feridex？ Ⅳ, Berlex Laboratories）,Fe r u c a r b o t ra n （ Re s ov i s t？, B aye r H e a l t h c a re ） ,Ferumoxtran-10 （AMI-227 or Code-7227, Combidex？, AMAG Pharma; Sinerem？, Guerbet）, NC100150（Clariscan？, Nycomed,） and （VSOP C184, Ferropharm）have been designed and clinically tested as magneticresonance contrast agents. However, until nowResovist？ is current available in only a few countries.The other four agents have been stopped for furtherdevelopment or withdrawn from the market. AnotherSPIO agent Ferumoxytol （Feraheme） is approved forthe treatment of iron deficiency in adult chronic kidneydisease patients. Ferumoxytol is comprised of ironoxide particles surrounded by a carbohydrate coat, andit is being explored as a potential imaging approach forevaluating lymph nodes and certain liver tumors.
