CAJ | 학술논문

目的：探讨小干扰RNA（siRNA）沉默EZH2表达对人膀胱癌细胞增殖、侵袭和迁移能力，以及对凋亡的影响。方法构建靶向EZH2基因的siRNA质粒并转染至人膀胱癌细胞中（干扰组），采用RT-PCR法检测EZH2mRNA的表达情况，利用噻唑蓝（MTT）、Transwell细胞小室及划痕实验检测转染后细胞增殖、侵袭和转移能力的变化，通过流式细胞术实验观察转染后细胞的凋亡情况。结果转染EZH2基因的siRNA质粒后（分干扰1组，干扰2组，干扰3组），干扰各组EZH2mRNA表达较阴性对照组有明显抑制（P＜0．05），且干扰2组效果最好，因此选干扰2组作后期实验；在转染48h后，干扰2组生长抑制率达37．9％，较阴性对照组受到明显抑制（P＜0．05）；划痕实验处理24h后，干扰2组细胞的迁移距离较阴性对照组明显降低（P＜0．01）；干扰2组细胞侵袭能力较阴性对照组下降了67％，差异有统计学意义（P＜0．01）；转染48h后，干扰2组早、晚期凋亡率分别为22．80％和3．60％，较阴性对照组增加，且以早期明显，差异有统计学意义（P＜0．01）。结论EZH2基因沉默能有效抑制人膀胱癌细胞的增殖、侵袭和迁移能力，并促进其凋亡，为深入研究膀胱癌的基因治疗提供理论依据。
목적：탐토소간우RNA（siRNA）침묵EZH2표체대인방광암세포증식、침습화천이능력，이급대조망적영향。방법구건파향EZH2기인적siRNA질립병전염지인방광암세포중（간우조），채용RT-PCR법검측EZH2mRNA적표체정황，이용새서람（MTT）、Transwell세포소실급화흔실험검측전염후세포증식、침습화전이능력적변화，통과류식세포술실험관찰전염후세포적조망정황。결과전염EZH2기인적siRNA질립후（분간우1조，간우2조，간우3조），간우각조EZH2mRNA표체교음성대조조유명현억제（P＜0．05），차간우2조효과최호，인차선간우2조작후기실험；재전염48h후，간우2조생장억제솔체37．9％，교음성대조조수도명현억제（P＜0．05）；화흔실험처리24h후，간우2조세포적천이거리교음성대조조명현강저（P＜0．01）；간우2조세포침습능력교음성대조조하강료67％，차이유통계학의의（P＜0．01）；전염48h후，간우2조조、만기조망솔분별위22．80％화3．60％，교음성대조조증가，차이조기명현，차이유통계학의의（P＜0．01）。결론EZH2기인침묵능유효억제인방광암세포적증식、침습화천이능력，병촉진기조망，위심입연구방광암적기인치료제공이론의거。
Objective To investigate the effect of EZH2 knockdown on cell proliferation ,invasion ,migration and apoptosis in hu-man bladder cancer cell line by small interfering RNAs(siRNA) .Methods The siRNA-expressing plasmid targeting EZH2 gene was constructed and transfected into T24 cells .RT-PCR was used to detect the EZH2 gene′s expression at the level of mRNA ;pro-liferation ,invasion and migration of T24 cells were examed in vivo by MTT ,wound healing assay and Transwell chamber migration assay .Finally ,Annexin V-FITC/PI flow cytometric analysis was performed for cell apoptosis .Results The siRNA-expressing plas-mid targeting EZH2 gene successfully inhibited EZH2 gene’s expression in T24 cells .The expression of mRNA was significantly inhibited compared with negative control groups (P<0 .05) .After the transfection of the plasmid 48 hours ,the growth inhibition rate was 37 .9% ,which was higher than the negative control group(P<0 .05) .24 hours after wound healing ,the migration distance of transfected group cell was(1 .37 ± 0 .12) ,which was lower than the negative control group(P<0 .01) .Compared with the nega-tive control group ,invasion capability of EZH2-siRNA group was dropped by 67% (P<0 .01) .48 hours after transfection ,the early and secondary apoptosis rate of T 24 cells were 22 .80% and 3 .60% respectively ,which were higher than the negative contral group (P<0 .01) .Conclusion The siRNA interference EZH2 can significantly inhibit cell proliferation ,invasion and migration of T24 cells ,meanwhile promote its apoptosis .It provides a theoretical basis for further study of bladder cancer gene therapy .