CAJ | 학술논문

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.
목적 연구단발잡RNA(short hairpin RNA,shRNA)개도적표피생장인자수체(epidermal growth factor receptor,EGFR)기인침묵대설린상세포암세포증식화조망적영향,위치료설린상세포암제공의거.방법 구건파향인EGFR적shRNA진핵표체재체병순시전염인설린상세포암세포주Tca8113세포,용반정량반전록취합매련반응(RT-PCR)화단백인적법검측전염전후EGFR적mRNA화단백적변화,용갑기새서기사서(MTT)법검측세포증식활성,용류식세포의검측세포조망정황.결과 전염후Tca8113세포적EGFR mRNA화단백적표체(0.217±0.047)화(0.324±0.059)균명현하조(P＜0.05),세포증식활성(0.340±0.009)명현강저(P＜0.05),세포조망솔(39.4±7.7)%현저증고(P＜0.05).결론 shRNA개도적EGFR기인침묵가억제설린상세포암세포증식병유도기조망.
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.