FHIT基因转染对肝癌细胞系HepG2的作用

Role of transfection of FHIT gene in human hepatocellular carci noma cell line HepG2

目的:探讨外源性FH IT基因的表达对肝癌细胞株HepG2的影响.方法:用脂质体介导法将pcDNA3.1-FH IT及空载体pcDNA3.1(+)导入肝癌细胞系HepG2中,G418选择培养,经免疫组化和W estern b lotting鉴定其过表达.用流式细胞仪、普通光镜、透射电镜观察FH IT基因的表达对HepG2细胞凋亡和增殖的影响.结果:获得FH IT基因稳定表达的肝癌细胞株HepG2-FH IT.转染FH IT基因的HepG2细胞的生长速率明显下降,电镜下超微结构发生了凋亡早期改变,流式分析细胞的生长周期可见S/G2期阻滞.结论:外源性FH IT基因在体外通过诱导其凋亡及细胞周期俘获作用可有效抑制HepG2生长增殖.

AIM： To study the role of transfection of FHIT gene into a human liver cancer cell line HepG2. METHODS： FHIT gene was transfected into human liver cancer cell line HepG2 by lipofectamine. After selection with G418, resistant colonies were obtained. The over-expression of FHIT was detected by immunochemistry and Western blotting methods. Apoptosis and cell cycle were analyzed by flow cytometry （FCM）. The morphological changes were observed by light and electron microscope. RESULTS： Stable expression of FHIT protein in transfected HepG2 cells was obtained and the growth rate of these transfected HepG2 was inhibited. Early apoptosis was observed under transmission electron microscope and flow cytometry showed that the progression of cell cycle was arrested from S phase to G2 phase. CONCLUSION： The results indicate that FHIT gene can inhibit the growth and proliferation of HepG2 by inducing its apoptosis and cell cycle arrest in vitro.