Wang Kunpeng, Ma Long, Tang Jingyuan, Yu Qiu, Shen Yang, Wei Yunfei, Zhu Chen, Deng Zhonglei, Zhang Wei
Department of Urology, Lianyungang Clinical College of Nanjing Medical University, Lianyungang, 222061, China.
Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China.
J Cancer. 2020 Jan 14;11(6):1468-1477. doi: 10.7150/jca.35710. eCollection 2020.
The purpose of our study is to elucidate the expression of lncRNA00518 (lnc00518) in the bladder cancer, and its potential mechanism in regulating the development of bladder cancer. The expression of lnc00518 in bladder cancer tissues and cells was examined by qRT-PCR. Correlation between lnc00518 expression with clinicopathologic characteristics and prognosis of bladder cancer patients was analyzed. effects of lnc00518 on the cellular behaviors of bladder cancer cells were explored. Moreover, effect of lnc00518 was evaluated by subcutaneous tumorigenesis in nude mice. The possible miRNA targets of lnc00518 were predicted by bioinformatics and further confirmed by dual-luciferase reporter gene assay, RIP and rescue experiments. Lnc00518 was highly expressed in bladder cancer tissues and cells. Lnc00518 expression was correlated with TNM staging and histological grade of bladder cancer. Besides, the overall survival was lower in bladder cancer patients with high expression of lnc00518 relative to those with low expression. Overexpression of lnc00518 enhanced proliferative, invasive, migratory potentials and clonality, but shortened G0/G1 phase of bladder cancer cells. Lnc00518 knockdown obtained the opposite trends. experiments revealed that lnc00518 knockdown inhibited subcutaneous tumorigenesis in nude mice. QRT-PCR results indicated that lnc00518 expression was negatively correlated with miRNA-101 expression in bladder cancer cells. Through dual-luciferase reporter gene assay and RIP, we confirmed the binding between lnc00518 and miRNA-101. Furthermore, EZH2 was verified to be the target of miRNA-101. MiRNA-101 knockdown reversed the inhibitory roles of lnc00518 knockdown in proliferative, migratory and invasive potentials of bladder cancer cells. Lnc00518 is highly expressed in bladder cancer and can be served as a predictor of poor prognosis. Lnc00518 promotes the proliferative, invasive and migratory potentials of bladder cancer by upregulating EZH2 competitively binding to miRNA-101.
我们研究的目的是阐明lncRNA00518（lnc00518）在膀胱癌中的表达及其调控膀胱癌发展的潜在机制。通过qRT-PCR检测lnc00518在膀胱癌组织和细胞中的表达。分析lnc00518表达与膀胱癌患者临床病理特征及预后的相关性。探讨lnc00518对膀胱癌细胞细胞行为的影响。此外，通过裸鼠皮下成瘤实验评估lnc00518的作用。通过生物信息学预测lnc00518可能的miRNA靶点，并通过双荧光素酶报告基因检测、RIP和拯救实验进一步证实。lnc00518在膀胱癌组织和细胞中高表达。lnc00518表达与膀胱癌的TNM分期和组织学分级相关。此外，lnc00518高表达的膀胱癌患者总生存期低于低表达患者。lnc00518的过表达增强了膀胱癌细胞的增殖、侵袭、迁移能力和克隆性，但缩短了G0/G1期。lnc00518敲低则得到相反的结果。实验表明lnc00518敲低抑制了裸鼠皮下成瘤。qRT-PCR结果表明lnc00518表达与膀胱癌细胞中miRNA-101表达呈负相关。通过双荧光素酶报告基因检测和RIP，我们证实了lnc00518与miRNA-101之间的结合。此外，EZH2被证实是miRNA-101的靶点。miRNA-101敲低逆转了lnc00518敲低对膀胱癌细胞增殖、迁移和侵袭能力的抑制作用。lnc00518在膀胱癌中高表达，可作为预后不良的预测指标。lnc00518通过竞争性结合miRNA-101上调EZH2，促进膀胱癌细胞的增殖、侵袭和迁移能力。
