Liu Yunfu, Xu Xin, Xu Xianglai, Li Shiqi, Liang Zhen, Hu Zhenghui, Wu Jian, Zhu Yi, Jin Xiaodong, Wang Xiao, Lin Yiwei, Chen Hong, Mao Yeqing, Luo Jindan, Zheng Xiangyi, Xie Liping
Department of Urology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China.
Oncol Lett. 2017 Nov;14(5):5121-5128. doi: 10.3892/ol.2017.6865. Epub 2017 Sep 1.
MicroRNAs (miRNAs) are small non-coding RNAs that affect various biological processes by altering the expression of a target gene. An miRNA microarray analysis has previously revealed a significant decrease in miR-193a-3p levels in prostate cancer tissues compared with that in their benign prostate hyperplasia counterparts. However, the role of miR-193a-3p has yet to be elucidated. In the present study, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to evaluate the expression levels of miR-193a-3p in two human prostate cancer cell lines. Forced overexpression of miR-193a-3p was established by transfecting mimics into DU-145 and PC3 cell lines. Cell proliferation and the cell cycle were assessed using a cell viability assay, flow cytometry and a colony formation assay. In addition, the target gene of miR-193a-3p was determined by a luciferase assay, RT-qPCR and western blot analysis. The regulation of the cell cycle by miR-193a-3p was also evaluated by western blotting. The results demonstrated that miR-193a-3p expression levels were lower in prostate cancer cell lines as compared with the RWPE normal prostate epithelium cell line. Subsequent gain-of-function studies revealed that stable miR-193a-3p transfection inhibited cell viability, proliferation and colony formation, and induced G phase arrest in prostate cancer cells. A luciferase assay and western blot analysis identified cyclin D1 () as a direct target gene of miR-193a-3p. In addition, the forced expression of was able to counter the inhibitory effects of miR-193a-3p transfection in the prostate cancer cells. In summary, the results suggest that miR-193a-3p may inhibit the viability, proliferation and survival of prostate cancer cells by regulating the expression profile of , and that miR-193a-3p may be a novel therapeutic biomarker for prostate cancer.
微小RNA(miRNA)是一类小的非编码RNA,通过改变靶基因的表达来影响各种生物学过程。先前的一项miRNA微阵列分析显示,与良性前列腺增生组织相比,前列腺癌组织中miR-193a-3p水平显著降低。然而,miR-193a-3p的作用尚未阐明。在本研究中,采用逆转录定量聚合酶链反应(RT-qPCR)来评估miR-193a-3p在两种人前列腺癌细胞系中的表达水平。通过将模拟物转染到DU-145和PC3细胞系中,实现了miR-193a-3p的强制过表达。使用细胞活力测定、流式细胞术和集落形成测定来评估细胞增殖和细胞周期。此外,通过荧光素酶测定、RT-qPCR和蛋白质印迹分析来确定miR-193a-3p的靶基因。还通过蛋白质印迹评估了miR-193a-3p对细胞周期的调控。结果表明,与RWPE正常前列腺上皮细胞系相比,前列腺癌细胞系中miR-193a-3p表达水平较低。随后的功能获得性研究表明,稳定转染miR-193a-3p可抑制前列腺癌细胞的活力、增殖和集落形成,并诱导G期停滞。荧光素酶测定和蛋白质印迹分析确定细胞周期蛋白D1()为miR-193a-3p的直接靶基因。此外,的强制表达能够抵消miR-193a-3p转染对前列腺癌细胞的抑制作用。总之,结果表明miR-193a-3p可能通过调节的表达谱来抑制前列腺癌细胞的活力、增殖和存活,并且miR-193a-3p可能是前列腺癌的一种新型治疗生物标志物。