siRNA沉默TGFBR2基因对HepG2细胞增殖的影响  

Significance of siRNA-mediated TGFBR2 gene silencing on HepG2 cell proliferation

作　　者：程林[1] 邓武坚[1] 蒋小峰[1] 于琨[1] 陈德[1] 

机构地区：[1]广州医科大学附属第二医院肝胆外科,510260

出　　处：《实用医学杂志》2014年第14期2200-2203,共4页The Journal of Practical Medicine

基　　金：广东省科技计划项目(编号:2010B060900088);广州市科技和信息化局项目(编号:201300000170)

摘　　要：目的:通过siRNA干扰技术研究TGFBR2基因沉默对HepG2细胞增殖的影响。方法:设计3种针对TGFBR2基因的siRNA片段,瞬时转染进HepG2细胞中;挑选出沉默效率最高的siRNA片段,并将对应的DNA序列插入到pEGFP-N3质粒中,再将质粒转染到HepG2细胞中,检测TGFBR2蛋白表达水平。用TGF-β1刺激siRNA干扰后的HepG2细胞,对比干扰前后细胞增殖的变化。结果:在3种siRNA片段中,siRNA-1的沉默效率最高。用5 ng/mL的TGF-β1刺激HepG2细胞,与blank组和siRNA-NC组比较,siRNA-1组细胞增殖加快(P<0.05),但仍低于未加TGF-β1因子的正常HepG2细胞(P<0.05)。结论:通过siRNA技术沉默TGFBR2基因后,可减弱TGF-β1信号传导通路对HepG2肝癌细胞株的抑制作用,导致肿瘤细胞的增殖加快。Objective To value the significance of TGFBR2 gene in mediating HepG2 cell proliferation by RNA interference technology. Methods Three kinds of siRNAs targeting TGFBR2 gene were designed, synthesized and transfected into HepG2 cells via lipofetamine2000. Among three kinds of siRNAs, only the one with the most interference efficacy was selected and the correspondent DNA sequence was inserted into plasmid pEGFP-N3. Then the recombinant plasmid of siRNA-pEGFP-N3 was transfected into HepG2 cell and western blot was used to detect the protein level of TGFBR2. Then, TGF-β1 was used to stimulate HepG2 cells with or without siRNA interference and proliferation of HepG2 cells was observed. Results Among these three siRNAs, siRN-1 appeared to be the most effective. After stimulated by 5ng/mL TGF-β1, proliferation of HepG2 cells showed a marked increase in siRNA-1 group compared with blank and siRNA-NC groups （P〈0.05）. For all that, the proliferation rate was still lower than that in normal HepG2 cell group without TGF-β1 stimulation. Conclusion By silencing TGFBR2 gene, inhibition of TGF-β1 signaling pathway to HepG2 cells could be decreased, thereby enhancing the cell proliferation.

关 键 词：癌 干细胞 Ⅱ型转化生长因子-β受体 HEPG2细胞 HEPG2 

分 类 号：R735.7[医药卫生—肿瘤]