RNA干扰MEX3A基因对膀胱癌细胞增殖和凋亡的影响

发布时间：2018-05-09 15:10

本文选题：膀胱癌 + MEXA基因　；参考：《解剖学报》2017年06期

【摘要】：目的探讨通过慢病毒介导的RNA干扰技术抑制MEX3A基因表达对膀胱癌细胞增殖和凋亡的影响。方法应用GV115载体构建针对MEX3A基因的shRNA慢病毒载体,转染包装细胞293T产生慢病毒颗粒,收集并滴度测定后转染5637膀胱癌细胞,实验组转染MEX3A-shRNA慢病毒,对照组转染阴性对照慢病毒;实时定量PCR(Real-time PCR)检测MEX3A基因敲减效率;慢病毒转染3 d后,使用Celigo连续细胞计数5d检测细胞生长;慢病毒转染5d后,进行膜联蛋白V(Annexin V)染色并用流式细胞术检测细胞凋亡。结果成功构建MEX3AshRNA慢病毒载体以及稳定抑制MEX3A表达的细胞株,MEX3A基因敲减效率达到74%;Celigo细胞计数检测显示,与对照组相比,实验组5637细胞的增殖速率受到显著抑制;流式细胞术检测显示,实验组发生凋亡的5637细胞显著增加。结论 MEX3A基因促进膀胱癌细胞的增殖,抑制膀胱癌细胞的凋亡。
[Abstract]:Objective to investigate the effect of lentivirus-mediated RNA interference on the proliferation and apoptosis of bladder cancer cells. Methods the shRNA lentivirus vector targeting MEX3A gene was constructed with GV115 vector. Lentivirus particles were produced by transfection of packaging cells 293T. After collecting and titer determination, 5637 bladder cancer cells were transfected with lentivirus. The experimental group transfected MEX3A-shRNA lentivirus and the control group transfected negative control lentivirus. MEX3A gene knockout efficiency was detected by real-time quantitative PCR(Real-time PCR, cell growth was detected by Celigo continuous cell count for 5 days after lentivirus transfection for 3 days, and cell apoptosis was detected by flow cytometry with V(Annexin V staining after 5 days of lentivirus transfection. Results the successful construction of MEX3AshRNA lentivirus vector and the knockdown efficiency of MEX3A gene reached 74%. Compared with the control group, the proliferation rate of 5637 cells in the experimental group was significantly inhibited. Flow cytometry showed that the number of apoptotic 5637 cells in the experimental group was significantly increased. Conclusion MEX3A gene can promote the proliferation of bladder cancer cells and inhibit the apoptosis of bladder cancer cells.
【作者单位】：中国医科大学附属盛京医院超声科;
【基金】：国家自然科学基金(81371552)
【分类号】：R737.14

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