Soochow University, Soochow, Jiangsu, 215000, China; Department of Urology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, 121000, China.
Crit Rev Eukaryot Gene Expr. 2022;32(8):81-94. doi: 10.1615/CritRevEukaryotGeneExpr.2022043175.
Bladder cancer is a common urological tumor, and its development process is complicated. Many long non-coding RNAs (lncRNAs), including PCAT19, have a regulatory role in cell development and gene expression, and is widely involved in the occurrence and development of a variety of cancers. We studied the changes in cell function through MTT assay, flow cytometer, transwell assay, etc. Several molecular assays are employed to explore these molecular mechanistic aspects, such as luciferase reporter, reverse transcription quantitative polymerase chain reaction, and Western blot. In addition, we also constructed a mouse bladder cancer xenograft model to verify the function of the PCAT19/miR-335-5p/IER2 signal axis in vivo. PCAT19 is upregulated in bladder cancer tissues, while miR-335-5p is downregulated, and the expression of PCAT19 and miR-335-5p is negatively correlated. The survival rate of bladder cancer patients in the PCAT19 upregulated group or miR-335-5p downregulated group was lower. After overexpression of PCAT19, the amount of miR-335-5p in cell is decreased, and the IER2 expression is increased. It can significantly promote the vitality, proliferation, migration and invasion capabilities of bladder cancer cells, and significantly inhibit cell apoptosis. Studies on molecular mechanisms show that PCAT19 can regulate the miR-335-5p/IER2 signal axis both in vivo and in vitro. PCAT19, which is upregulated in bladder cancer tissue, interacts with miR-335-5p to regulate the expression of downstream target gene IER2, forming a complete PCAT19/miR-335-5p/IER2 signal regulation axis, and ultimately affect the malignant progression of bladder cancer both in vivo and in vitro.
膀胱癌是一种常见的泌尿系统肿瘤,其发展过程较为复杂。许多长链非编码 RNA(lncRNA),包括 PCAT19,在细胞发育和基因表达中具有调节作用,广泛参与多种癌症的发生和发展。我们通过 MTT 检测、流式细胞术、Transwell 检测等方法研究细胞功能的变化,运用几种分子检测方法探讨这些分子机制,如荧光素酶报告基因检测、逆转录定量聚合酶链式反应(RT-qPCR)和 Western blot 等。此外,我们还构建了小鼠膀胱癌移植瘤模型,以验证 PCAT19/miR-335-5p/IER2 信号轴在体内的功能。PCAT19 在膀胱癌组织中上调,而 miR-335-5p 下调,且 PCAT19 和 miR-335-5p 的表达呈负相关。PCAT19 上调组或 miR-335-5p 下调组膀胱癌患者的生存率较低。过表达 PCAT19 后,细胞中 miR-335-5p 的含量减少,IER2 的表达增加。它能显著促进膀胱癌细胞的活力、增殖、迁移和侵袭能力,显著抑制细胞凋亡。分子机制研究表明,PCAT19 可在体内和体外调节 miR-335-5p/IER2 信号轴。在膀胱癌组织中上调的 PCAT19 与 miR-335-5p 相互作用,调节下游靶基因 IER2 的表达,形成完整的 PCAT19/miR-335-5p/IER2 信号调控轴,最终影响膀胱癌的体内外恶性进展。