CAJ | 학술논문

目的观察小分子干扰RNA(siRNA)对胃癌MGC803细胞E2F-1基因表达及其生物学行为的影响.方法将实验组(E2F-1-siRNA)和阴性对照组(negative control-siRNA)转染进MGC803细胞,48 h后采用实时定量聚合酶链反应(PCR)及Western blot技术分别检测E2F-1 mRNA及蛋白的表达,噻唑蓝(MTT)比色法检测MGC803细胞增殖的变化,流式细胞仪检测MGC803细胞周期变化.结果 E2F-1-siRNA有效抑制胃癌细胞E2F-1 mRNA和蛋白的表达并且影响MGC803细胞的增殖,与阴性对照组及未转染组细胞比较mRNA降低了84.8%和85.3%(F=14.35,P＜0.05),蛋白降低T77.2%和79.6%,MGC803细胞增殖分别抑制了69.0%和81.6%,差异有统计学意义(F=170.18,P＜0.05);同时E2F-1-siRNA促使更多MGC803细胞DNA含量聚集于G2/M期附近,G2/M期DNA含量比例为(54.98±3.46)%,与阴性对照组(44.70±3.82)%和未转染组(31.47±2.12)%比较,差异有统计学意义(F=88.90,P＜0.05).结论 E2F-1-siRNA可以有效抑制人胃癌MGC803细胞E2F-1 mRNA及蛋白的表达,使G1 期细胞的DNA含量下调,细胞分裂停滞在G2/M期附近,抑制胃癌细胞的增殖.
목적 관찰소분자간우RNA(siRNA)대위암MGC803세포E2F-1기인표체급기생물학행위적영향.방법 장실험조(E2F-1-siRNA)화음성대조조(negative control-siRNA)전염진MGC803세포,48 h후채용실시정량취합매련반응(PCR)급Western blot기술분별검측E2F-1 mRNA급단백적표체,새서람(MTT)비색법검측MGC803세포증식적변화,류식세포의검측MGC803세포주기변화.결과 E2F-1-siRNA유효억제위암세포E2F-1 mRNA화단백적표체병차영향MGC803세포적증식,여음성대조조급미전염조세포비교mRNA강저료84.8%화85.3%(F=14.35,P＜0.05),단백강저T77.2%화79.6%,MGC803세포증식분별억제료69.0%화81.6%,차이유통계학의의(F=170.18,P＜0.05);동시E2F-1-siRNA촉사경다MGC803세포DNA함량취집우G2/M기부근,G2/M기DNA함량비례위(54.98±3.46)%,여음성대조조(44.70±3.82)%화미전염조(31.47±2.12)%비교,차이유통계학의의(F=88.90,P＜0.05).결론 E2F-1-siRNA가이유효억제인위암MGC803세포E2F-1 mRNA급단백적표체,사G1 기세포적DNA함량하조,세포분렬정체재G2/M기부근,억제위암세포적증식.
Objective To investigate the effects of RNA interference(RNAi)targeting E2F-1 on biological behaviors of gastric cancer cell line MGC803.Methods E2F-1-siRNA or negative control-siRNA was transfected into gastric cancer cell line MGC803 cells.Forty-eight h later,the expression of E2F-1 mRNA and protein in gastric cancer cell fine MGC803 was detected by real-time quantitative PCR and Western blot respectivelv.Cell cycle was examined by flow cytometry,and the proliferative rate of MGC803 cells tegted by MTT assay.Results E2F-1-siRNA effectively inhibited the expression of E2F-1 mRNA and Drotein and influenced proliferation of MGC803 cells.As compared with negative control and nontmsfected groups,the expression levels of E2F-1 mRNA and protein were decreased by 84.8%,85.3%(P＜0.05),and 77.2%,79.6%,and proliferation of MGC803 cells was suppressed by 69.0%and 81.6%(P＜0.05)in E2F-1-siRNA transfection group,respectively.E2F-1-siRNA promoted accumulation of more DNA of MGC803 cells at G2/M phase,and the rate of DNA content in G2/M phase in E2F-1-siRNA trailsfection group was(54.98±3.46)%,which was significantly higher than(43.07±3.82)% in negative control group,and(31.47±2.12)%in non-transfection group(P＜0.05).Conclusion E2F-1 gene repression by RNAi can decrease the DNA content in G1 phase of MGC803 cells,which can arrest the cells in G2/M phase and inhibit the proliferation of MGC803 cells.