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目的 研究短发卡RNA(short hairpin RNA,shRNA)介导的表皮生长因子受体(epidermal growth factor receptor,EGFR)基因沉默对舌鳞状细胞癌细胞增殖和凋亡的影响,为治疗舌鳞状细胞癌提供依据.方法 构建靶向人EGFR的shRNA真核表达载体并瞬时转染人舌鳞状细胞癌细胞株Tca8113细胞,用半定量反转录聚合酶链反应(RT-PCR)和蛋白印迹法检测转染前后EGFR的mRNA和蛋白的变化,用甲基噻唑基四唑(MTT)法检测细胞增殖活性,用流式细胞仪检测细胞凋亡情况.结果 转染后Tca8113细胞的EGFR mRNA和蛋白的表达(0.217±0.047)和(0.324±0.059)均明显下调(P＜0.05),细胞增殖活性(0.340±0.009)明显降低(P＜0.05),细胞凋亡率(39.4±7.7)%显著增高(P＜0.05).结论 shRNA介导的EGFR基因沉默可抑制舌鳞状细胞癌细胞增殖并诱导其凋亡.

Objective To investigate the effects of epidermal growth factor receptor (EGFR)gene silencing mediated by short hairpin RNA ( shRNA ) on proliferation and apoptosis of human tongue carcinoma cells. Methods shRNA eukaryotic expression vector targeting the specific sequence of human EGFR gene was constructed and termed shEGFR. The control vector targeting the unrelated sequence was also constructed and termed shNC. The vectors were transiently transfected into Tca8113 cells of human tongue squamous cell carcinoma by LipofectamineTM2000, respectively. The mRNA and protein levels of EGFR in Tca8113 cells were detected by reverse transcriptase polymerase chain reaction (RT-PCR)and Western blotting. The cell proliferation of Tca8113 cells was evaluated by methyl thiazolyl tetrazolium(MTT) assay. The apoptosis of Tca8113 cells was assessed by flow cytometry. Results EGFR expression in Tca8113 cells transfected with shEGFR were obviously decreased at mRNA level (81.6% )and protein level (72.0%) (P＜0.05) 48 h after transfection of shEGFR compared with untransfected Tca8113 cells. The proliferation activity of Tca8113 cells transfected with shEGFR was significantly lower than that of Tca8113 cells transfected with shNC and untransfected Tca8113 cells (P＜0. 05). The early apoptotic rate of Tca8113 cells transfected with shEGFR was significantly higher than that of Tca8113 cells transfected with shNC and untransfected Tca8113 cells [ (39.4±7.7)%,(4.3±1.2)%,(2.5±0.9)%.P＜0.05] 48 h after transfection of shEGFR.Conclusions EGFR gene silencing mediated bv shRNA may inhibit cell proliferation and induce apoptosis in human tongue carcinoma cells.