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人肝细胞生长因子基因表达对CCl_4损伤的人肝细胞及大鼠肝星状细胞株的影响及其机制

Effects of human hepatocyte growth factor gene on CCl_4-injured human hepatocytes and rat hepatic stellate cell line and their mechanisms
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摘要 目的:探讨人肝细胞生长因子(hHGF)基因的表达对CCl4损伤的人肝细胞及大鼠肝星状细胞株(CFSC-2G)生物学效应的影响及其对CCl4损伤的人正常肝细胞株(HL-7702)的保护作用,进一步探讨凋亡产生的机制。方法:将获得的稳定转染HL-7702细胞克隆,分为4组,Ⅰ组为转染PCI-neo-hHGF实验组,Ⅱ组为转染PCI-neo空载体对照组,Ⅲ组为仅加入脂质体的对照组,Ⅳ组为空白对照组。采用MTT法测定细胞增殖活性。采用Westernblotting测定转染PCI-neo-hHGF的HL-7702细胞和CFSC-2G细胞中hHGF、caspase-3、caspase-8、caspase-9蛋白质的表达。结果:HL-7702细胞转染PCI-neo-HGF并培养48h后,细胞的增殖活性明显高于PCI-neo空载体对照组、脂质体转染对照组和空白对照组HL-7702细胞(P<0.01),而且随着转染PCI-neo-HGF剂量增加,细胞的增殖活性有一定的增长趋势,但各组间比较差异无显著性(P>0.05);PCI-neo-HGF转染的HL-7702细胞caspase-3蛋白质表达量较CCl4损伤的HL-7702细胞和PCI-neo转染的HL-7702细胞明显降低,未检测到caspase-8和caspase-9蛋白质的表达;PCI-neo-HGF转染的CFSC-2G细胞caspase-3蛋白质表达量较CCl4诱导的CFSC-2G增加,caspase-9蛋白质表达也呈阳性。结论:PCI-neo-HGF可促进体外培养的HL-7702细胞的生长增殖,能够抑制CCl4损伤的HL-7702细胞的凋亡,促进大鼠CFSC-2G细胞的凋亡,其作用机制可能与上调caspase-3和caspase-9蛋白质表达有关。 Objective To explore the biological effects of human hepatocyte growth factor （hHGF） gene expression on CCl4-injured human hepatocytes and rat hepatic stellate cell line and the protective effect of hHGF gene expression on CCl4-injured human normal liver cell line （HL-7702）,and explore the mechanism of apoptosis.Methods The stably transfected HL-7702 cell clones were divided into four groups：PCI-neo-hHGF group,PCI-neo empty vector group,liposome control group,and blank control group.The proliferative activity of cells was determined by MTT method.The protein expressions of hHGF,caspase-3,caspase-8,and caspase-9 were measured using Western blotting in PCI-neo-hHGF transfected HL-7702 cells and CFSC-2G cells.Results After the HL-7702 cells were transfected with PCI-neo-HGF and cultivated for 48 h,the cell proliferation activity was significantly higher than those in PCI-neo,liposome and control groups （P0.01）;and with the increasing of transfected dose of PCI-neo-HGF,the proliferation activity of the cells had a trend of increasing,but there was no significant difference between various groups（P0.05）.The protein expression of caspase-3 in PCI-neo-HGF transfected HL-7702 cells was decreased significantly compared with those in CCl4-injured HL-7702 cells and PCI-neo transfected HL-7702 cells;the protein expressions of caspase-8 and caspase-9 were not detected;the protein expression of caspase-3 in PCI-neo-HGF transfected CFSC-2G cells was increased compared with those in CCl4-induced CFSC-2G cells,the protein expression of caspase-9 was also positive.Conclusion PCI-neo-HGF can promote the growth and proliferation of HL-7702 cells,and inhibit the apoptosis of CCl4-injured HL-7702 cells,and promote the apoptosis of rat CFSC-2G cells in vitro.Its mechanism may be related to the up-regulation of the protein expressions of caspase-3 and caspase -9.
作者 李玉香 张凯宇 张一宁 王峰 姜艳芳 牛俊奇
机构地区 吉林大学第一医院感染症科
出处 《吉林大学学报（医学版）》 CAS CSCD 北大核心 2010年第3期505-509,共5页 Journal of Jilin University：Medicine Edition
基金 吉林省科技厅科研基金资助课题(20060417-2)
关键词 人肝细胞生长因子 CFSC-2G细胞 细胞凋亡 CASPASE-3 CASPASE-8 CASPASE-9 human hepatocyte growth factor CFSC-2G cells apoptosis caspase-3 caspase-8 caspase-9
分类号 Q78 [生物学—分子生物学]
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参考文献13

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吉林大学学报（医学版）
2010年 第3期

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