CAJ | 학술논문

目的观察小干扰RNA(siRNA)沉默表皮生长因子受体(EGFR)对人肾癌细胞株(ACHN)细胞增殖、迁移及侵袭能力的影响.方法化学合成针对EGFR的siRNA,实验分3组,通过脂质体转染法转染进ACHN肾癌细胞中,利用Western blot技术检测细胞中EGFR蛋白的敲除效率,通过锥虫蓝拒染法测定细胞生长曲线,用噻唑蓝(MTT)比色分析法、细胞划痕实验、Transwell分别检测细胞的增殖、迁移及侵袭能力.结果特异性siRNA可明显抑制EGFR的表达；生长曲线提示,RNA干扰(RNAi)组细胞在24、48、72、96 h增殖抑制率分别为15.6％、38.4％、64.1％、68.0％,较空白对照组及阴性对照组生长明显减慢(P＜0.05)；24h后RNAi组划痕宽度为(109.97±7.50) μm,空白对照组及阴性对照组的划痕基本愈合(P＜0.05)；RNAi组穿膜细胞数[(153±13)个],分别与空白对照组[(320±20)个]、阴性对照组[(300±16)个]比较,穿膜细胞明显减少(P＜0.05).结论 siRNA抑制EGFR表达后,肾癌细胞的增殖、迁移及侵袭能力显著降低.
목적 관찰소간우RNA(siRNA)침묵표피생장인자수체(EGFR)대인신암세포주(ACHN)세포증식、천이급침습능력적영향.방법 화학합성침대EGFR적siRNA,실험분3조,통과지질체전염법전염진ACHN신암세포중,이용Western blot기술검측세포중EGFR단백적고제효솔,통과추충람거염법측정세포생장곡선,용새서람(MTT)비색분석법、세포화흔실험、Transwell분별검측세포적증식、천이급침습능력.결과 특이성siRNA가명현억제EGFR적표체；생장곡선제시,RNA간우(RNAi)조세포재24、48、72、96 h증식억제솔분별위15.6％、38.4％、64.1％、68.0％,교공백대조조급음성대조조생장명현감만(P＜0.05)；24h후RNAi조화흔관도위(109.97±7.50) μm,공백대조조급음성대조조적화흔기본유합(P＜0.05)；RNAi조천막세포수[(153±13)개],분별여공백대조조[(320±20)개]、음성대조조[(300±16)개]비교,천막세포명현감소(P＜0.05).결론 siRNA억제EGFR표체후,신암세포적증식、천이급침습능력현저강저.
Objective To investigate the effects of small interfering RNA (siRNA) silencing epidermal growth factor receptor (EGFR) gene on the ability of proliferation,migration and invasion of the renal cell carcinoma cell line renal cancer cell line (ACHN).Methods ACHN cells were transfected with siRNA targeting human EGFR by liposome transfection reagent,and three experimental groups were set up.The expression level of EGFR was detected by using Western blotting.The cell growth curves were determined by trypan blue staining.The proliferation inhibition of ACHN cells was measured by methyl thiazolyl tetrazolium (MTT) assay.The migration of ACHN cells was detected by using wound healing assay,and the ability of invasion was examined by Transwell assay.Results The expression of EGFR gene could be suppressed obviously by sequence-specific shRNA targeting EGFR gene.The growth curve revealed the cell proliferation inhibition rate in RNAi group was 15.6％,38.4％,64.1％,and 68.0％ at 24,48,72,and 96 h respectively,which was significantly reduced as compared with the blank control group and negative group (P ＜0.05).In RNAi group cell scratch was (109.97 7.5) μm after 24 h,and cell scratches in blank control group and negative group were basically healed (P ＜ 0.05).The number of migarating cells in RNAi group was (153 ± 13),which was significantly reduced as compared with the blank control group [(320 20)] and negative control group [(300 ± 16)] (P ＜ 0.05).Conclusion siRNA targeting human EGFR could specially suppress the expression of EGFR gene in renal cell carcinoma cells,which could significantly reduce the capability of proliferation,migration and invasion of the renal cell carcinoma cells.