CAJ | 학술논문

目的观察核糖核酸(RNA)干扰对TES基因表达及肿瘤细胞增殖的影响.方法将TES基因特异性的小分子干扰RNA(siRNA)转入肿瘤细胞株Ishikawa,实时荧光定量聚合酶链反应(Real-time PCR)技术检测TES mRNA的表达,Western blot法进行蛋白定量,噻唑蓝(MTT)比色法检测细胞增殖,研究其对TES基因的表达及细胞增殖的影响.结果 RNA干扰后12、24、48、72 h,实验组中TES mRNA表达量分别为0.675、0.024、0.324、0.916,较各对照组明显降低,差异均有统计学意义(P＜0.05)；而在不同时间点比较对照A组、对照B组及空白组TES mRNA的表达差异无统计学意义(P＞0.05).实验组中TES mRNA的表达在24 h时最低.RNA干扰后Ishikawa细胞中TES蛋白的表达与TES mRNA水平变化一致,转染后12 ～48 h内,实验组TES蛋白的表达量明显降低分别为65.2、3.2、25.6、33.0,差异均有统计学意义(P＜0.05)；TES蛋白表达在24 h时达最低水平.针对TES基因的RNA干扰后Ishikawa细胞的增殖活性明显增强(P＜0.05).结论针对TES基因的RNA干扰后,TES mRNA和蛋白水平均呈显著下降趋势,同时,Ishikawa细胞的增殖活性明显升高,证实了TES基因在子宫内膜癌中所发挥的抑癌基因的功能.
목적 관찰핵당핵산(RNA)간우대TES기인표체급종류세포증식적영향.방법 장TES기인특이성적소분자간우RNA(siRNA)전입종류세포주Ishikawa,실시형광정량취합매련반응(Real-time PCR)기술검측TES mRNA적표체,Western blot법진행단백정량,새서람(MTT)비색법검측세포증식,연구기대TES기인적표체급세포증식적영향.결과 RNA간우후12、24、48、72 h,실험조중TES mRNA표체량분별위0.675、0.024、0.324、0.916,교각대조조명현강저,차이균유통계학의의(P＜0.05)；이재불동시간점비교대조A조、대조B조급공백조TES mRNA적표체차이무통계학의의(P＞0.05).실험조중TES mRNA적표체재24 h시최저.RNA간우후Ishikawa세포중TES단백적표체여TES mRNA수평변화일치,전염후12 ～48 h내,실험조TES단백적표체량명현강저분별위65.2、3.2、25.6、33.0,차이균유통계학의의(P＜0.05)；TES단백표체재24 h시체최저수평.침대TES기인적RNA간우후Ishikawa세포적증식활성명현증강(P＜0.05).결론 침대TES기인적RNA간우후,TES mRNA화단백수평균정현저하강추세,동시,Ishikawa세포적증식활성명현승고,증실료TES기인재자궁내막암중소발휘적억암기인적공능.
Objective To observe impact of RNA interference of the TES gene expression and tumor cell proliferation.Methods Small interfering RNA (siRNA) specific-targeting TES gene was transfected into endometrial cancer cell line Ishikawa,studying its effects on TES expression with quantitative Real-time polymerase chain reaction (Real-time PCR) and western blotting,cellular proliferation was determined by methyl thiazol tetrazolium (MTT) assay.Results At different time point (12,24,48,72 h),the TES mRNA expression in experimental group were 0.675,0.024,0.324,0.916,significantly lower compared with control (P ＜ 0.05) ; there was no statistically significant difference between the control groups (P ＞ 0.05).MRNA level of TES reached the lowest at 24 h in the experimental group.TES protein in Ishikawa cells was consistent with TES mRNA levels change after RNA interference:12-48 h after transfection,the experimental group TES protein expression were 65.2,3.2,25.6,33.0,was significantly reduced,and the differences were statistically significant (P ＜ 0.05) ; TES protein expression was also the lowest at 24 h.After transfection with siRNA,the proliferation activity of Ishikawa cells was enormously enhanced (P ＜ 0.05).Conclusion After RNA interference,TES mRNA and protein levels showed a significant downward trend,proliferation of Ishikawa cells also was significantly enhanced,confirmed that TES gene functioned as a tumor suppressor in endometrial cancer.