摘要
目的观察小分子干扰RNA(siRNA)沉默CD147基因在肝癌细胞HepG2中的表达,探讨CD147基因对肝癌细胞生长、凋亡的影响。方法根据CD147 cDNA序列设计具有短发夹结构的两条DNA序列,与载体pSilencerTM 4.1-CMV neo构建重组表达载体,鉴定后转染至HepG2细胞,Western blotting法检测抑制效果,MTT法检测细胞增殖情况,流式细胞术检测肿瘤细胞凋亡情况。结果成功构建了针对CD147基因表达的干扰质粒,有效抑制了HepG2细胞增殖,促进了HepG2细胞凋亡。结论 CD147靶向RNA干扰重组表达载体为肝癌的基因治疗提供了可能。
Objective To evaluate small interfering RNAs（siRNAs） for downregulation of the CD147 expression in HepG2.Methods According to the gene sequence of CD147 in Genbank,one DNA sequence containing a small hairpin structure were designed and synthesized.The complement form was cloned into vector pSilencerTM 4.1-CMV neo,and the recombinant plasmid was transfected into HepG2.The inhibitory effects were detected by Western blotting,the cell proliferation was detected by MTT,and the cell apoptosis was measured by flow cytometry.Results The recombinant plasmid was successful cloned and inhibited CD147 protein expression specifically.The cell growth in siRNA group was significantly inhibited after 48 h in cell growth medium.Conclusion Successful cloning of the recombinant plasmid may help to discover new gene therapies for hepatoma.
出处
《中国药业》
CAS
2011年第1期7-8,共2页
China Pharmaceuticals
