Department of Urology, Wuhan Third Hospital, Tongren Hospital of Wuhan University, No. 216 Guanshan Avenue, Hongshan District, Wuhan, 430014, Hubei, China.
Mol Biotechnol. 2022 Nov;64(11):1259-1269. doi: 10.1007/s12033-022-00489-x. Epub 2022 May 13.
This study is to clarify the effect of the long non-coding RNA ArfGAP with RhoGAP domain, ankyrin repeat and PH domain 1-antisense RNA 1 (ARAP1-AS1)/microRNA (miR)-3918/kinesin family member 20A (KIF20A) on bladder cancer cell function. ARAP1-AS1, miR-3918, and KIF20A expression levels in bladder cancer cells were determined using quantitative reverse transcription-polymerase chain reaction. The effects of ARAP1-AS1, miR-3918, and KIF20A on bladder cell activity, proliferation, apoptosis, and in vivo growth were examined using the cell counting kit-8, colony formation, caspase-3 activity, and xenograft tumor growth assays, respectively, in nude mice. The binding relationships among ARAP1-AS1, miR-3918, and KIF20A were analyzed using luciferase and RNA immunoprecipitation assays. ARAP1-AS1 and KIF20A were overexpressed in bladder cancer, while miR-3918 was underexpressed. The downregulation of ARAP1-AS1 or KIF20A expression significantly inhibited the viability and proliferation of cancer cells and promoted apoptosis, whereas low expression of miR-3918 or high expression of ARAP1-AS1/KIF20A showed the opposite effect. miR-3918 was sponged by ARAP1-AS1, and targeted KIF20A. In addition, miR-3918 expression was inversely correlated with ARAP1-AS1 and KIF20a expression levels in bladder cancer tissues. In addition, the rescue experiment showed that interference with miR-3918 could reverse the effect of low ARAP1-AS1 or KIF20A expression on bladder cancer cell malignancy. ARAP1-AS1 facilitates the malignant behavior of bladder cancer cells via the regulation of KIF20A expression by sponging miR-3918.
本研究旨在阐明长链非编码 RNA ArfGAP 域、ankyrin 重复和 PH 域 1-反义 RNA 1(ARAP1-AS1)/microRNA(miR)-3918/驱动蛋白家族成员 20A(KIF20A)对膀胱癌细胞功能的影响。采用实时定量逆转录聚合酶链反应检测膀胱癌细胞中 ARAP1-AS1、miR-3918 和 KIF20A 的表达水平。分别采用细胞计数试剂盒-8、集落形成、半胱天冬酶-3 活性和裸鼠异种移植肿瘤生长试验检测 ARAP1-AS1、miR-3918 和 KIF20A 对膀胱细胞活性、增殖、凋亡和体内生长的影响。采用荧光素酶和 RNA 免疫沉淀试验分析 ARAP1-AS1、miR-3918 和 KIF20A 之间的结合关系。ARAP1-AS1 和 KIF20A 在膀胱癌中过度表达,而 miR-3918 表达下调。下调 ARAP1-AS1 或 KIF20A 的表达显著抑制癌细胞的活力和增殖,并促进凋亡,而 miR-3918 低表达或 ARAP1-AS1/KIF20A 高表达则表现出相反的效果。miR-3918 被 ARAP1-AS1 海绵吸收,并靶向 KIF20A。此外,miR-3918 的表达与膀胱癌组织中 ARAP1-AS1 和 KIF20a 表达水平呈负相关。此外,挽救实验表明,干扰 miR-3918 可以逆转低 ARAP1-AS1 或 KIF20A 表达对膀胱癌细胞恶性行为的影响。ARAP1-AS1 通过海绵吸附 miR-3918 调节 KIF20A 的表达,促进膀胱癌细胞的恶性行为。