昆明医科大学学报2016，Vol.37Issue(8)：19-23,5.
慢病毒介导的FHIT基因过表达调控人肝癌细胞株生长实验研究
Effects of FHIT Overexpression Mediated by Slow Virus on the growth of Hepatoma Lines in Vitro
摘要
Abstract
Objective To observe overexpressional effects of FHIT gene on proliferation,apoptosis,invasion and cell cycle of human hepatocellular carcinoma cells including HepG2,Hep3B and Huh7.Method The pLVX-IRES-FHIT slow virus recombinant vector was constructed and transfected into HepG2,Hep3B and Huh7 cells.We detected the infection of cell proliferation,apotosis,invasion and cell cycle of these cells with overexpression of FHIT and analyzed the relationship of biological effects and time gradient by MTT,Annexin V/PI using cell cycle detection technology and Transwell experiment.Results According to DNA sequencing,the pLVX-IRES-FHIT slow virus recombinant vector was constructed successfully.(1) MTI tests showed that proliferation activity of HepG2,Hep3B and Huh7 with pLVX-IRES-FHIT transfection was reduced significantly in 48 hours and the inhibition was stable after 72 hours.The inhabitation in the control group was not significant (P＜ 0.05).AnnexinV/PI double staining method showed that no significant difference of apoptosis level was seen after 48h among three hepatoma cells (P＞0.05).The order of inhibitional proliferation activity from high to low was HepG2,Hep3B and Huh7;(2) Transwell tests showed that invasive activity of HepG2,Hep3B and Huh7 with pLVX-IRES-FHIT transfection was inhibited (P＜0.05).The control group was not sinificantly inhibited (P＜ 0.05).The order of invasive activity was HepG2 ＞ Hep3B ＞ Huh7;(3) Cell cycle test showed that the number of cells in G0/G1 stage increased whereas decreased in stage S.Hep3B and Huh7 cells in the control group were not significantly inhibited (P＞0.05).The influence of FHIT over expression to these cells was that cell cycle stopped in S stage,and sequence of cell number in G0/G1 was HepG2 ＜Hep3B ＜Huh7.Conclusion A positive correlation was found between FHIT expression and cellular differentiation on hepatoma ceils.FHIT gene may be a potential gene therapy for human hepatocellular carcinoma.关键词
FHIT/慢病毒载体/增殖/凋亡/细胞侵袭性/细胞周期Key words
FHIT gene/Slow virus recombinant vector/Proliferation/Apoptosis/Invasion/Cell cycle分类
医药卫生引用本文复制引用
戈佳云,杨筱燕,李晓,吴雪松,施智甜,赵松凌,张文瀚,王琳,魏东..慢病毒介导的FHIT基因过表达调控人肝癌细胞株生长实验研究[J].昆明医科大学学报,2016,37(8):19-23,5.基金项目
国家自然科学基金资助项目(81061434) （81061434）
