摘要
目的:研究小干扰RNA(small interfering RNA,siRNA)抑制高迁移率族蛋白1(HMGB1)基因的表达对人前列腺癌细胞PC-3增殖的影响。方法:构建真核表达载体Pgenesil-1/HMGB1 siRNA,在脂质体Lipofectamine 2000的介导下转染前列腺癌PC-3细胞株,通过RT-PCR和Western Blot检测HMGB1的mRNA及蛋白质的变化;流式细胞仪检测细胞周期;噻唑蓝(MTT)检测细胞生长曲线。观察转染后PC-3细胞HMGB1基因表达和体外增殖活性的变化。结果:成功构建siRNA表达载体Pgenesil-1/HMGB1 siRNA,所获表达载体转染可使PC-3细胞HMGB1 mRNA和蛋白表达水平显著降低(P<0.05),并能有效抑制PC-3增殖活性(P<0.05)。结论:应用siRNA干扰技术能有效的抑制HMGB1基因的表达,同时也可有效抑制癌细胞的体外增殖活性,为肿瘤的生物学治疗提供新思路。
Objective： To study the effect of specific inhibition of high mobility group Box-1 gene expression by small interfering RNAs （siRNA） on the proliferation of human prostate carcinoma cell line PC-3. Methods： We constructed and identified an eukaryotic expression vector carrying hu,nan Pgenesil-1/HMGB1 siRNA.The vector was transfeeted into PC-3 cells by Lipofectamine 2000 and the stable clones were selected with G418. The HMGB1 expression in PC-3 cells was detected by RT-PCR and Western blot before and after transfeetion. Flow eytometry was employed to detect the percentage of cells in each phase of the cell cycle. The in vitro proliferation activity was examined by MTT analysis. Resuits： The recombinant plasmid Pgenesil-1/HMGB1 siRNA was suceessfully constructed. The introduction of Pgenesil-1/ HMGB1 siRNA efficiently and specifically inhibited the expression of the HMGB1 gene aceording to the resuhs of RTPCR and Western blot, with a significant difference between the siRNA-transfeeted group and the group without siRNA transfection （P〈0.05）. Cell proliferation in the Pgenesil-1/HMGB1 siRNA group was significantly inhibited （P〈0.05）. Conclusion： siRNA targeting HMGB1 mRNA can specifically suppress HMGB1 gene expression and can effectively inhibit the proliferation of PC-3 cells, providing a potential new method for cancer biotherapy.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2008年第6期339-343,共5页
Chinese Journal of Clinical Oncology
基金
国家973计划基金资助(编号:2002CB513100)~~
