CAJ | 학술논문

目的:探讨shRNA干扰沉默PLC ε基因对人膀胱癌T24细胞生长的影响.方法:体外构建PLCε基因的shRNA的重组质粒,Lipofectamilie2000介导法转染T24细胞,采用RT-PCR方法检测特异性shRNA的重组质粒对PLCε基因沉默效果,转染后采用MTT法检测shRNA的重组质粒对细胞增殖的作用,免疫细胞化学染色法检测T24细胞PCNA的表达,电镜观察细胞超微结构的改变.结果:shRNA的重组质粒能有效下调PLCε基因的表达,抑制率为78.01%,与对照组比较其差异有统计学意义(P<0.01);细胞增殖活性受到明显抑制,与对照组比较其差异有统计学意义(P<0.01);细胞内PCNA含量明显低于空白对照组和阴性质粒组,其差异有统计学意义(P<0.01);细胞形态改变产生凋亡小体.结论:PLCε基因有望成为应用RNAi技术探索治疗膀胱癌潜在的靶基因.
목적:탐토shRNA간우침묵PLC ε기인대인방광암T24세포생장적영향.방법:체외구건PLCε기인적shRNA적중조질립,Lipofectamilie2000개도법전염T24세포,채용RT-PCR방법검측특이성shRNA적중조질립대PLCε기인침묵효과,전염후채용MTT법검측shRNA적중조질립대세포증식적작용,면역세포화학염색법검측T24세포PCNA적표체,전경관찰세포초미결구적개변.결과:shRNA적중조질립능유효하조PLCε기인적표체,억제솔위78.01%,여대조조비교기차이유통계학의의(P<0.01);세포증식활성수도명현억제,여대조조비교기차이유통계학의의(P<0.01);세포내PCNA함량명현저우공백대조조화음성질립조,기차이유통계학의의(P<0.01);세포형태개변산생조망소체.결론:PLCε기인유망성위응용RNAi기술탐색치료방광암잠재적파기인.
Objective:To investigate whether PLC ε gene down regulation by RNA interference (RNAi) leads to inhibition of proliferation in human bladder carcinoma T24 cell. Methods: The shRNA recombinant plasmids targeting to PLC ε gene was constructed and transfected into bladder carcinoma T24 cell with Lipofectamine 2000. RT-PCR was used to monitor the validity of specific s h R N A in down regulation of PLC ε . Then MTT assay was performed for detecting cell proliferation, the changes of PCNA were analyzed by immunocytochemical method,Electron microscope was used to observe the morphological changes. Results: The specific PLC ε shRNA was confirmed to be efficient in silencing PLC e expression. PLC ε gene down regulation by the shRNA recombinant plasmids inhibition cell proliferation rate about 78.01%, They were significantly different from that of control group (P<0.01; After transfected the specific recombinant plasmids,PCNA expression was significantly decreased,They were significantly different from that of control group (P<0.01;morphological changes produce apoptotic body. Conclusion: PLCε is likely to be potential molecular target for bladder carcinoma in gene therapy by RNAi.