Nie Yong-Li, Tang Shi-Min, Peng Fang, Xu Tao, Mao Yong-Rong
Department of Oncology, Sinopharm Hanjiang Hospital, Danjiangkou 442700.
Department of Functionology, Yangtze University, Jingzhou 434023.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Nov;36(6):637-641. doi: 10.12047/j.cjap.6031.2020.133.
To investigate the effects of miRNA-191 on the proliferation, migration and invasion of prostate cancer, and to explore its mechanism. The expression levels of miRNA-191 in four human prostate cancer cell lines (PC-3, DU-145, LNCa P, 22RU1) and human normal prostate cell line RWPE-2 were detected, and prostate cancer cell line PC-3 was selected as the experimental object. PC-3 cells were divided into three groups: blank control group (no transfection), miRNA-191 NC group (PC-3 cells transfected with Inhibitor NC) and miRNA-191 Inhibitor group (PC-3 cells transfected with miRNA-191 Inhibitor), and each group was provided with three multiple pores. The expression levels of miRNA-191 and PLCD1 were detected by RT-PCR. The cell proliferation was detected by CCK8 assay. Scratch test and invasive test were used to detect cell migration and invasive ability. Through Targetscan target gene prediction website, PLCD1 was screened as the target protein of miRNA-191, and verified by double luciferase target experiment.Western blot assay was used to detect the expression of PLCD1 in cells of each group. Compared with RWPE-2 cells, the expression level of miRNA-191 in human prostate cancer cells was significantly higher ( ＜0.05), and the expression level of miRNA191 in PC-3 was significantly higher than that in other three cell lines (＜0.05). After inhibiting the expression of miRNA-191, the expression levels of PLCD1 was significantly higher while PC-3 cells' proliferation ability was inhibited, and their migration and invasion ability were significantly lower than those of blank control group and miRNA-191 NC group (＜ 0.05). The results of double luciferase reporter gene assay showed that PLCD1 gene was a target gene of miRNA-191. miRNA-191 promote the proliferation, migration and invasion of prostate cancer PC-3 cells by targeting PLCD1.
探讨miRNA-191对前列腺癌细胞增殖、迁移及侵袭的影响，并探究其作用机制。检测了4种人前列腺癌细胞系（PC-3、DU-145、LNCa P、22RU1）及人正常前列腺细胞系RWPE-2中miRNA-191的表达水平，选取前列腺癌细胞系PC-3作为实验对象。将PC-3细胞分为3组：空白对照组（未转染）、miRNA-191 NC组（转染Inhibitor NC的PC-3细胞）和miRNA-191抑制剂组（转染miRNA-191抑制剂的PC-3细胞），每组设置3个复孔。采用RT-PCR检测miRNA-191和PLCD1的表达水平。采用CCK8法检测细胞增殖情况。划痕试验和侵袭试验检测细胞迁移和侵袭能力。通过Targetscan靶基因预测网站筛选出PLCD1为miRNA-191的靶蛋白，并通过双荧光素酶靶标实验进行验证。采用Western blot法检测各组细胞中PLCD1的表达。与RWPE-2细胞相比，人前列腺癌细胞中miRNA-191的表达水平显著升高（＜0.05），PC-3中miRNA-191的表达水平显著高于其他3种细胞系（＜0.05）。抑制miRNA-191表达后，PLCD1的表达水平显著升高，同时PC-3细胞的增殖能力受到抑制，其迁移和侵袭能力显著低于空白对照组和miRNA-191 NC组（＜0.05）。双荧光素酶报告基因检测结果显示，PLCD1基因是miRNA-191的靶基因。miRNA-191通过靶向PLCD1促进前列腺癌PC-3细胞的增殖、迁移及侵袭。
