Wan Pei, Chen Zhilin, Huang Minzhi, Jiang Huiming, Wu Huajun, Zhong Kaihua, Ding Guodong, Wang Bing
Department of Urology, Meizhou People's Hospital (Huangtang Hospital), Meizhou, China.
Department of Urology, Shangrao Municipal Hospital, Shangrao, China.
Transl Androl Urol. 2021 Nov;10(11):4262-4274. doi: 10.21037/tau-21-941.
MicroRNAs (miRs) are endogenous, single-stranded, noncoding RNAs that are involved in various physiological processes, and the development and the progression of various types of cancer. Specifically, the role of miR-200a-3p has been implicated in various types of cancer in contributing to a diverse array of cancer types has been previously reported. The present study aimed to investigate the expression levels of miR-200a-3p in human bladder cancer, as well as its potential role in disease pathogenesis.
Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of has-mir-200a-3p and tumor necrosis factor α () induced protein 3 () in tumor tissues and cell lines. Dual-luciferase reporter assay and combination with the expression intervention of hsa-mir-200a-3p and in bladder cancer cell lines to clarify the binding relationship between hsa-mir-200a-3p and .After the expression intervention of hsa-mir-200a-3p and in bladder cancer cells, the changes of cell proliferation, cell apoptosis, cell cycle, wound-healing ability and migration ability were detected by CCK8, flow cytometry, wound-healing and Transwell methods. Xenograft transplantation model was performed subcutaneously in nude mice by implantation of J82 and T24 cells, and then the bladder cancer growth curve was calculated from mice exposed to has-mir-200a-3p minic or minic-NC.
Bladder cancer tissues demonstrated significantly upregulated miR-200a-3p expression levels. Moreover, increased miR-200a-3p expression was significantly associated with distant metastasis and advanced stage. In addition, compared with the miR-control (Ctr) group, miR-200a-3p overexpression promoted bladder cancer cell proliferation, migration, invasion, cell cycle, and release of inflammatory cytokines, but inhibited cell apoptosis. Mechanistically, was identified as a target gene of miR-200a-3p in bladder cancer cell lines. Moreover, compared with the miR-Ctr group, the miR-200a-3p overexpression group exhibited significantly promoted tumor growth , and overexpression blocked the promoting effect of miR-200a-3p on bladder cancer.
The results of the present study indicated that miR-200a-3p might serve act as an oncogene in human bladder cancer by targeting a novel the gene gene; therefore, miR-200a-3p and might serve could serve as novel therapeutic targets for bladder cancer.
微小RNA(miRs)是内源性单链非编码RNA,参与多种生理过程以及各类癌症的发生和发展。具体而言,miR - 200a - 3p在多种癌症中的作用先前已有报道。本研究旨在探讨miR - 200a - 3p在人膀胱癌中的表达水平及其在疾病发病机制中的潜在作用。
采用逆转录聚合酶链反应(RT - PCR)和蛋白质免疫印迹法检测肿瘤组织和细胞系中has - mir - 200a - 3p和肿瘤坏死因子α(TNFα)诱导蛋白3(TNFAIP3)的表达。通过双荧光素酶报告基因检测并结合在膀胱癌细胞系中对hsa - mir - 200a - 3p和TNFAIP3进行表达干预,以阐明hsa - mir - 200a - 3p与TNFAIP3之间的结合关系。在膀胱癌细胞中对hsa - mir - 200a - 3p和TNFAIP3进行表达干预后,采用CCK8法、流式细胞术、伤口愈合实验和Transwell法检测细胞增殖、细胞凋亡、细胞周期、伤口愈合能力和迁移能力的变化。通过将J82和T24细胞皮下接种到裸鼠体内建立异种移植模型,然后计算接受hsa - mir - 200a - 3p模拟物或模拟物阴性对照的小鼠的膀胱癌生长曲线。
膀胱癌组织中miR - 200a - 3p表达水平显著上调。此外,miR - 200a - 3p表达增加与远处转移和晚期显著相关。另外,与miR - 对照(Ctr)组相比,miR - 200a - 3p过表达促进膀胱癌细胞增殖、迁移、侵袭、细胞周期进程以及炎性细胞因子释放,但抑制细胞凋亡。机制上,TNFAIP3被确定为膀胱癌细胞系中miR - 200a - 3p的靶基因。此外,与miR - Ctr组相比,miR - 200a - 3p过表达组肿瘤生长显著加快,而TNFAIP3过表达可阻断miR - 200a - 3p对膀胱癌的促进作用。
本研究结果表明,miR - 200a - 3p可能通过靶向新基因TNFAIP3在人膀胱癌中发挥癌基因作用;因此,miR - 200a - 3p和TNFAIP3可能成为膀胱癌新的治疗靶点。