miR-345靶向调控 TGM1抑制膀胱癌的初步研究

目的:探讨mi R-345调控TGM1表达影响膀胱癌的分子生物学机制。方法:首先,采用RT-qPCR检测T24和RT4细胞中mi R-345、TGM1的表达;再采用mi RNA-NC、mi R-345 mimic、NC inhibitor、mi R-345 inhibitor、control si RNA、si TGM1和pc-DNA3.1/TGM1等转染膀胱癌细胞;然后,采用MTT实验检测细胞增殖,Transwell实验检测细胞侵袭,流式细胞仪检测细胞凋亡,双荧光报告酶检测mi R-345的靶基因;最后,采用Western blot检测TGM1在细胞中的表达。结果:mi R-345在T24和RT4细胞中表达低于正常细胞(P0.05)。mi R-345过表达时,T24和RT4细胞的增殖侵袭能力减弱,细胞凋亡率上升;mi R-345表达沉默时,细胞增殖和侵袭能力增强,细胞凋亡率下降。双荧光报告基因检测结果显示TGM1为mi R-345的靶基因,mi R-345过表达抑制TGM1的表达(P0.05);mi R-345表达沉默时则表达上调(P0.05)。当TGM1表达沉默时,T24和RT4细胞的增殖和侵袭能力减弱,细胞凋亡率上升;TGM1过表达时该细胞的增殖和侵袭能力增强,细胞凋亡率下降。结论:mi R-345通过下调靶基因TGM1的表达,抑制膀胱癌细胞的增殖、侵袭并促进细胞凋亡。

miR-345 Inhibits the Bladder Cancer by Inhibiting Target Gene TGM1

ABSTRACT Objective: To explore the molecular mechanism of miR-345-regulated bladder cancer through targeting TGM1. Methods:First, the expression of miR-345 and TGM1 in T24 or RT4 were detected by RT-qPCR. Then, the miRNA-NC, miR-345 mimic, NC-inhibitor, miR-345 inhibitor, control siRNA, siTGM1 and pc-DNA3.1/TGM1were transfected into the bladder cancer cells. The proliferation and invasion was detected by MTT assay and Transwell assay, respectively. The flow cytometry was used to detected the apoptosis of the cells. The target gene of miR-345 was detected by Double fluorescent reporter. Finally, the protein expression of TGM1 in cells was detected by Western blot. Results:The expression of miR-345 in T24 and RT4 cells were lower than in normal cells. The abilities of proliferation and invasion of T24 and RT4 cells were decreased and the apoptosis rates were increased when miR-345 was overexpressed. While the expression of miR-345 was silenced, the abilities of proliferation and invasion were enhanced, and the apoptosis rates were decreased. We found that TGM1 was the target gene of miR-345 by Dual fluorescent reportor, and the expression of TGM1 was inhibited by the overexpression of miR-345 (P<0.05), the expression was up-regulated when the expression of miR-345 was silenced (P<0.05). The abilities of proliferation and invasion of T24 and RT4 cells were decreased, and the apoptosis rates were increased. The abilities of proliferation and invasion were increased, and the apoptosis rates were decreased when the TGM1 was overexpressed. Conclusion:miR-345 inhibits the proliferation, invasion and apoptosis of bladder cancer cells by decreasing the expression of target gene TGM1.