山西医科大学学报2011，Vol.42Issue(6)：443-446,4.DOI:10.3969/J.ISSN.1007-6611.2011.06.001
siRNA干扰HSP90α表达对人肝癌HepG2细胞影响的研究
Effect of HSP90α RNA interference on biological behaviour of liver cancer cell line HepG2
摘要
Abstract
Objeaive To establish a c:ell line stahly inhibiting the heat shock protein 90α( HSP90α )expression by siRNA interference and to explore the effect of HSP90α siRNA on proliferation and cell cycle of liver cancer cell line HepG2. Methocis The recombinant plasmid pSilencerHSP90 containing the 2lnt small interfering RNA of human HSP90α was subcloned, purified and identified by DNA sequence analysis. Then the recombinant plasmid pSilencerHSP90 was introduced into HepG2 cell by electroporation. After G418 selection, positive clones were isolated and cultured to form a cell line stably inhibiting HSP90α expression( siRNA interference group ). The non-transfected HepG2 cells were chosen as control group. The positive clones were identified by quantitative RT-PCR and Western blot. CCK-8 assay and flow cytometry( FCM ) were applied to determine the cell growth and cell cycle , respectively. Results The sequence of specific siRNA was correct by sequence analysis. The levels of HSP90a mRNA and protein expression were reduced in siRNA interference group. The CCK-8 assay results showed that the cell growth in siRNA interference group was significantly inhibited after 48 h compared with control group. The FCM results showed that there was an obvious G2/M phase arrest in siRNA interference group.Conclusion The cell lines stably low expressing HSP90α is succ:essfully established. The decreased expression of HSP90agene can inhihit the cell proliferation.and c:ause the cell cycle arrest. Successful cloning of the recombinant plasmid may help to find a new gene therapy for hepatoma.关键词
HSP90/RNA干扰/HepG2细胞Key words
HSP90 / RNA interference / HepG2 cells分类
医药卫生引用本文复制引用
王晓捷,尹强兵,马晓姣,邹飞,陈雪梅..siRNA干扰HSP90α表达对人肝癌HepG2细胞影响的研究[J].山西医科大学学报,2011,42(6):443-446,4.基金项目
国家自然科学基金资助项目(305000580,30971193) （305000580,30971193）
广东省自然科学基金资助项目(05300465)为肝癌的基因治疗提供理论基础. （05300465）
