环状 RNA 相互作用蛋白 RAPGEF5 通过海绵吸附 miR-582-3p 靶向 KIF3A 调控膀胱癌的增殖、迁移及侵袭。

CircRAPGEF5 sponges miR-582-3p and targets KIF3A to regulate bladder cancer cell proliferation, migration and invasion.

机构信息

Department of Urology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China.

出版信息

Int Immunopharmacol. 2024 Apr 20;131:111613. doi: 10.1016/j.intimp.2024.111613. Epub 2024 Mar 14.

DOI:10.1016/j.intimp.2024.111613

PMID:38489970

Abstract

BACKGROUND

Bladder cancer (BCa) is a common malignant disease with high recurrence and poor prognosis. Several circular RNAs (circRNAs) have been found to be associated with the malignant progression of bladder cancer (BCa). Here, the aim of this study was to investigate the expression, role and mechanism of circRAPGEF5 in BCa progression.

METHODS

Quantitative real-time PCR (qRT-PCR) and immunoblotting were used to detect gene and protein expression levels. In vitro functional studies were performed using CCK-8, colony formation, wound healing and Transwell assays, respectively, and a mouse xenograft tumor model was established to perform in vivo experiments. Bioinformatic predictions as well as luciferase reporter assays and RNA pull-down assays were used to probe circRAPGEF5-mediated competitive endogenous RNA (ceRNA) network.

RESULTS

CircRAPGEF5 was significantly overexpressed in BCa patients (p < 0.05), indicating a potential unsatisfactory prognosis. Functionally, knockdown of circRAPGEF5 inhibited the growth, migration and invasion of BCa cells in vitro (p < 0.05), as well as BCa growth in vivo (p < 0.05). Mechanistically, circRAPGEF5 acted as a sponge for miR-582-3p and targeted kinesin family member 3A (KIF3A). In addition, rescue experiments showed that inhibition of miR-582-3p or overexpression of KIF3A reversed the anticancer effects of circRAPGEF5 knockdown on BCa cells (p < 0.05).

CONCLUSION

Silencing circRAPGEF5 inhibits BCa proliferation, migration and invasion via the miR-582-3p/KIF3A axis, demonstrating a promising target for BCa-targeted therapy.

摘要

背景

膀胱癌（BCa）是一种常见的恶性疾病，具有高复发率和预后不良的特点。已经发现几种环状 RNA（circRNA）与膀胱癌（BCa）的恶性进展有关。本研究旨在探讨 circRAPGEF5 在 BCa 进展中的表达、作用和机制。

方法

采用定量实时 PCR（qRT-PCR）和免疫印迹法检测基因和蛋白表达水平。分别采用 CCK-8、集落形成、划痕愈合和 Transwell 实验进行体外功能研究，并建立小鼠异种移植肿瘤模型进行体内实验。通过生物信息学预测以及荧光素酶报告基因实验和 RNA 下拉实验，探讨 circRAPGEF5 介导的竞争性内源 RNA（ceRNA）网络。

结果

circRAPGEF5 在 BCa 患者中显著过表达（p<0.05），提示预后可能不佳。功能上，circRAPGEF5 的敲低抑制了 BCa 细胞在体外的生长、迁移和侵袭（p<0.05），以及体内的 BCa 生长（p<0.05）。机制上，circRAPGEF5 作为 miR-582-3p 的海绵，并靶向驱动蛋白家族成员 3A（KIF3A）。此外，挽救实验表明，抑制 miR-582-3p 或过表达 KIF3A 逆转了 circRAPGEF5 敲低对 BCa 细胞的抗癌作用（p<0.05）。