针对不同靶位点的siRNA对HepG2人肝肿瘤细胞株FOXO3a表达的抑制作用  

Inhibition of small interference RNA targeting FOXO3a on expression of FOXO3a in HepG2

作　　者：熊清芳[1] 杨永峰[1] 谢玉桃[2] 黄平[1] 钟艳丹[1] 冯晓宁[1] 

机构地区：[1]东南大学附属第二医院肝病科,南京210003 [2]中南大学湘雅医院传染科

出　　处：《中华临床医师杂志（电子版）》2013年第14期150-152,共3页Chinese Journal of Clinicians(Electronic Edition)

基　　金：湖南省自然科学基金(09JJ5031);中国肝炎基金(TQGB2011009);南京市医学重点科技发展项目(ZKX12039)

摘　　要：目的探讨针对FOXO3a的小干扰RNA(siRNA)对HepG2 FOXO3a的表达、细胞增殖的抑制及细胞凋亡的作用。方法利用RNA干扰(RNAi)效应设计针对FOXO3a的不同siRNA,构建表达载体质粒并转染HepG2细胞系。RT-PCR和Western Blot分别检测FOXO3a mRNA及蛋白的表达,细胞生长实验和流式细胞观察转染前后细胞增殖及细胞凋亡的变化。另设一无意义RNA载体质粒为阴性对照。结果P2、P3实验组,细胞核绿色荧光明显减淡,有细胞核的排出。特异性siRNA对FOXO3a mRNA及蛋白的表达具有抑制作用(P<0.05),细胞凋亡减少,增殖明显增加(P<0.05)。结论抑制FOXO3a基因的表达能够减少HepG2细胞的凋亡,并增加细胞的增殖。FOXO3a有望成为治疗肝癌的有效的靶基因。Objective To explore the effects of FOXO3a siRNAs on FOXO3a expression,cell cirele and apoptosis of HepG2 cells.Methods Three FOXO3a specific siRNAs were selected with the suppressing effect. RNAi expression vectors were constructed and transfected into HepG2 cells.RT-PCR and Western Blot were used to detect the expression of FOXO3a.The proliferation of the cells was measured by MTT test.The apoptosis of HepG2 cells was measured by flow cytometry.A nonsense small RNA was set as negative control. Results There were the export of FoxO3a to the cytoplasm from nucleus in P2,P3 group. When compared with the control groups,the FOXO3a-specific siRNAs inhibited the expression of FOXO3a in the protein and mRNA level, and suppressed the proliferation and induced the apoptosis of the HepG2 cells （P〈0.05）.Conclusion Knockdown expression of FOXO3a can decrease apoptosis and increase proliferation of HepG2 tumor cell,and FOXO3a gene can be a powerful target gene in treating liver cancer.

关 键 词：FOXO3A 肝肿瘤 RNA干扰 

分 类 号：R735.7[医药卫生—肿瘤]