重组腺病毒Canstatin对人肝癌HepG2细胞增殖影响的研究

Effects of Recombinant Adenovirus Canstatin on Proliferation of Human Hepatocellular Carcinoma HepG2 Cells

为研究重组腺病毒Canstatin感染人肝癌HepG2细胞及细胞转染后Canstatin在HepG2细胞中的表达,探讨Canstatin基因对人肝癌HepG2细胞生长增殖的影响。采用不同感染复数(MOI=10、40、80)的腺病毒Ad-Canstatin-GFP感染HepG2细胞,倒置荧光显微镜下观察细胞形态,流式检测感染效率,Real-Time PCR和Western blot法检测HepG2细胞中Canstatin mRNA和蛋白表达。CCK-8法检测细胞增殖能力的变化,Western blot法检测细胞中PCNA蛋白的表达。结果显示MOI为40时,72 h感染率可达到98.9%,HepG2细胞中Canstatin mRNA和蛋白表达均高于对照组和空载体组(P<0.05)。HepG2细胞感染Ad-Canstatin腺病毒后生长增殖受到抑制(P<0.05),且PCNA的表达低于对照组(P<0.05)。由此可知,Canstatin抑制人肝癌HepG2细胞的生长增殖有作用可能与影响PCNA的表达有关。

To study the transfection Canstatin expression in human HepG2 liver cancer cells infected by recombinant adenovirus Canstatin,and to explore Canstatin gene effects on proliferation of human hepatocellular carcinoma HepG2 cells.HepG2 cells were infected by different multiplicity （MOI=10,40,80） of adenovirus,which were labeled by GFP.Inverted fluorescence microscope was used to observe cell morphology, Flow cytometry to detect efficiency of infection,Real-Time PCR and Western blot method to examine HepG2 cells Canstatin mRNA and protein expression.CCK-8 method to masure the change of cell proliferation.The results showed that when the MOI of 40,72 h infection rate could reach 98.9%,In HepG2 cells,Canstatin mRNA and protein expression was higher than that of control group and empty carrier group （P〈0.05）.HepG2 cells proliferation was inhibited after infected with adenovirus Ad-Canstatin （P〈0.05）,and the expression of PCNA is lower than the control （P〈0.05）.Accordingly,Canstatin inhibits growth and proliferation of human hepatocellular carcinoma HepG2 cells may be related to affect the expression of PCNA.