载体介导shRNA对胃癌细胞株MKN45中C-met的抑制作用  

Inhibiting effect by vector-mediated shRNA on C-met in stomach cancer cell strain MKN45

作　　者：黄志盛 何海萍 梁锐彬 

机构地区：[1]广州市番禺区沙湾人民医院消化内科,511400

出　　处：《中国实用医药》2016年第5期26-27,共2页China Practical Medicine

摘　　要：目的探讨载体介导短发夹核糖核酸(shRNA)对胃癌细胞株MKN45中原癌基因C-met的抑制作用。方法 9株胃癌细胞株分为转染组、阴性对照组和空白对照组,各3侏。通过实时荧光定量逆转录-聚合酶链反应(RT-PCR)技术,观察转染24 h和48 h后,原癌基因C-met的信使核糖核酸(m RNA)的CT值的变化;通过免疫印迹试验(Western blot),观察转染后C-met的蛋白变化;通过四氮唑盐MTT比色法,计算细胞存活率。结果载体介导的C-met shRNA转染24 h和48 h后,RT-PCR结果显示,C-met的m RNA抑制率分别为56%和88%;与空白对照组比较,C-met的蛋白表达水平明显下降,转染后24 h[(0.812±0.105)VS(0.640±0.011)]、48 h[(0.811±0.103)VS(0.331±0.009)],差异均有统计学意义(P<0.05);MTT比色法显示,细胞存活率均有明显下降,转染后24 h为65.9%,48 h为59.6%。结论载体介导的C-met shRNA不仅可降低胃癌细胞株MKN45中原癌基因C-met的m RNA及其蛋白表达水平,而且对癌细胞的存活有明显的抑制作用。Objective To investigate inhibiting effect by vector-mediated short hairpin RNA（shRNA） on proto-oncogene C-met in stomach cancer cell strain MKN45. Methods A total of 9 stomach cancer cell strains were divided into transfection group, negative control group and blank control group, with 3 strains in each group. Real-time fluorescent quantitative reverse transcription-polymerase chain reaction（RT-PCR） was applied to observe changes of CT value of messenger ribonucleic acid（m RNA） in proto-oncogene C-met after 24 and 48 h of transfection. Western blot was used to observe changes of protein in C-met after transfection. Tetrazolium-based colorimetric assay（MTT） was applied to calculate cell survival rate. Results In 24 and 48 h after vector-mediated m RNA shRNA transfection, RT-PCR showed m RNA inhibition rates by C-met were respectively 56% and 88%. Comparing with the blank control group, C-met had obviously reduce protein expression level, as [（0.812±0.105） VS（0.640±0.011）] in 24 h after transfection and [（0.811±0.103） VS（0.331±0.009）] in 48 h after transfection. Their differences all had statistical significance（P〈0.05）, MTT showed obviously decreased cell survival rate, as 65.9% in 24 h after transfection and 59.6% in 48 h after transfection. Conclusion Vector-mediated C-met shRNA can not only reduce m RNA and its protein expression in proto-oncogene C-met of stomach cancer cell strain MKN45, and also show remarkable inhibiting effect for survival of cancer cell.

关 键 词：载体介导 胃癌 信使核糖核酸 原癌基因C-MET 

分 类 号：R735.2[医药卫生—肿瘤]