肝肠钙粘连蛋白对人肝癌细胞株Hep3B生长的影响  

The effects of LI-cadherin on the growth of Hep3B hepatocarcinoma cells and the related mechanisms

作　　者：范正军[1] 彭飞[1] 薛建峰[1] 程波[1] 刘艳杰[1] 

机构地区：[1]郑州大学第一附属医院肝胆外科,郑州450052

出　　处：《陕西医学杂志》2013年第2期147-150,共4页Shaanxi Medical Journal

摘　　要：目的:探讨肝肠钙粘连蛋白(LI-cadherin)对人肝癌细胞增值的影响,并初步研究其机制。方法:利用Lipofectamine脂质体转染法,将siRNA转染入Hep3B中,通过Western Bloting(蛋白质印迹)、MTT(四唑盐比色法)、Pl/Rnase染色-流式细胞分析法检测细胞中LI-cadherin蛋白及细胞周期相关蛋白CyclinD1表达量变化情况、LI-cadherin对肝癌细胞生长的影响并绘制细胞生长曲线及细胞周期分布。结果:空白对照组、空质粒转染组LI-cadherin的表达无明显差异(P>0.05),SiRNA转染组LI-cadherin的表达均低于空白对照组、空质粒转染组(P<0.05),SiRNA转染组与空白对照组、空质粒转染组相比,Cyclin D1蛋白表达明显升高;SiRNA转染组细胞生长速度在12h、24h和36h与空白对照组、空质粒转染组差异无统计学意义(P>0.05),在48h、72h和96h,则明显快于空白对照组和空质粒转染组(P<0.05);SiRNA转染组和空白对照组、空质粒转染组相比,S期及G2/M期百分比明显增多,G0/G1期百分比明显减少,细胞增殖指数(PI)较高,差异均具有统计学意义(P<0.05)。结论:成功建立了LI-cadherin-SiRNA-Hep3B细胞株,LI-cadher-in-SiRNA肝癌细胞株Hep3B中LI-cadherin蛋白表达降低,CyclinD1蛋白的表达增加。LI-cad-herin可抑制肝癌细胞株Hep3B的生长,其作用机制可能与其下调细胞周期蛋白Cyclin D1的表达水平及影响处于各周期细胞的百分含量的分布有关。Objective：To explore the effects and mechanism of LI-cadherin on the growth of human Hep- atocarcinoma Cells. Methods：SiRNA was transfected into Hep3B by Lipofectamine transfection method , and western bloting,MTT,PI/Rnase staining-Flow Cytometry（FCM） was used to detect the expression of Ll-cadherin and Cy- clinD1,the growth of human Hepatocarcinoma Cells and draw the cell growth curve, and detect the distribution of cell cycle in each group. Results：There was no significance in the expression levels of LI-cadherin between blank con- trol group and empty plasmid transfection group （P〉0.05）. The expression levels of LI-cadherin in the SiRNA transfection group was significantly lower than that in the blank control group and empty plasmid transfection group （P〈0.05）. The expression levels of Cyclin D1 in the SiRNA transfection group was significantly higher than that in the blank control group and empty plasmid transfection group（P〈0.05）. There was no significance in the levels of growth among blank control group, empty plasmid transfection group and SiRNA transfection group in 12h ,24h and 36h（P〈0.05）, the growth of SiRNA transfection group was significantly faster than that in the blank control group and empty plasmid transfection group in 48h,72h and 96h（P〈0.05）. Compared with blank control group and empty plasmid transfected group, the percentage of S phase and G2/M phase were increased significantly, G0/G1 phase was decreased significantly, cell proliferation index （PI） was more higher, and the differences were statistically sig- nificant（P〈0.05）. Conclusion：LI-cadherin-SiRNA-Hep3B cell line is constructed. The expression levels of LI-cad- herin is lower,and Cyclin D1 higher. LI-cadherin can inhibit the growth of the hepatoma cell line Hep3B, and the mechanisms ia probably related with its down-regulation the expression of Cyclin D1 levels and effect the percentage of cells in different cell cycle.

关 键 词：癌 肝细胞 细胞周期 生长 RNA干扰 细胞周期蛋白质类 

分 类 号：R735.7[医药卫生—肿瘤]