摘要
目的:探讨小RNA干扰(small interfering RNA,RNAi)对胃癌细胞株RegIα基因表达的抑制作用及对细胞增殖和凋亡的影响。方法:RT-PCR检测6株胃癌细胞的RegIα基因mRNA表达水平;体外构建RegIα基因小RNA干扰的重组质粒,分别稳定转染RegIα高表达细胞株MKN45和AGS,并设空载体作为对照组;应用RT-PCR和Western blot测定转染后细胞RegIα基因及其蛋白的表达水平;应用MTT法和流式细胞仪检测RegIα小RNA干扰对细胞增殖抑制率和凋亡的影响。结果:RT-PCR显示:与空载体组相比,转染RegIα小RNA干扰质粒后,MKN45和AGS细胞的RegIαmRNA明显被抑制;Western blot结果显示:MKN45和AGS细胞的RegIα蛋白表达水平分别降低至空载体组的(44±4)%和(25±4)%;MTT结果显示:RegIα小RNA干扰后MKN45和AGS细胞增殖均受到抑制,凋亡率分别为(12.96±0.50)%和(11.59±1.10)%,与空载体组比差异有统计学意义(P<0.05)。结论:小RNA干扰可有效抑制胃癌细胞株中RegIα基因的表达,具有抑制细胞增殖和诱导细胞凋亡的重要作用。
Objective: To investigate the effect of small interfering RNA on cell proliferation and apoptosis in gastric cancer cell lines with high expression of RegIα.Methods: Total RNA was isolated from six gastric cancer cell lines,and the expression of RegIα mRNA was detected by RT-PCR.RegIα RNAi expression vector was constructed and stably transfected into MKN45 and AGS cells with high RegIα expression,empty-vector was used as control.RegIα mRNA and protein expression was measured by RT-PCR and Western blot respe...
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2011年第1期57-63,共7页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省自然科学基金(Y206280)
浙江省教育厅科研项目(Y20070071)
关键词
RNA
小分子干扰
再生/遗传学
基因表达
胃肿瘤
遗传载体
质粒
细胞凋亡
细胞增殖
转染
RNA
small interfering
Regeneration/genet
Gene expression
Stomach neoplasms
Genetic vectors
Plasmids
Apoptosis
Cell proliferation
Transfection
