南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (1): 9-16.doi: 10.12122/j.issn.1673-4254.2024.01.02

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RBMX通过下调PKM2抑制膀胱癌细胞的增殖、迁移、侵袭和糖酵解

颜秋霞，曾 鹏，黄树强，谭翠钰，周秀琴，乔 静，赵晓英，冯 玲，朱振杰，张国志，胡 鸿，陈彩蓉   

  1. 广州医科大学附属第六医院/清远市人民医院生殖医学中心，泌尿外科，广东 清远 511518；广东省尿控及生殖医学创新工程技术研究中心，广东 清远 511518
  • 发布日期:2024-01-24

RBMX overexpression inhibits proliferation, migration, invasion and glycolysis of human bladder cancer cells by downregulating PKM2

YAN Qiuxia, ZENG Peng, HUANG Shuqiang, TAN Cuiyu, ZHOU Xiuqin, QIAO Jing, ZHAO Xiaoying, FENG Ling, ZHU Zhenjie, ZHANG Guozhi, HU Hong, CHEN Cairong   

  1. Center for Reproductive Medicine, Department of Urology, Sixth Affiliated Hospital of Guangzhou Medical University/ Qingyuan People's Hospital, Qingyuan 511518, China; Guangdong Engineering Research Center of Urinary Continence and Reproductive Medicine, Qingyuan 511518, China
  • Published:2024-01-24

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摘要： 目的 探讨X连锁RNA结合基序蛋白（RBMX）对膀胱癌细胞（1376细胞和UC-3细胞）的增殖、迁移、侵袭的影响以及其在糖酵解中的作用。方法 采用慢病毒表达系统和siRNA干扰技术，分别构建RBMX过表达和敲低的膀胱癌细胞模型（1376细胞和UC-3细胞）。采用RT-qPCR和Western blotting分别在mRNA水平和蛋白水平上检测细胞模型是否构建成功。通过EdU增殖实验和克隆形成实验检测过表达和敲低RBMX后细胞的生长和集落形成能力，同时通过Transwell实验分析过表达和敲低RBMX后对细胞迁移、侵袭能力的影响；随后，采用Western blotting检测过表达和敲低RBMX后糖酵解关键蛋白PKM1（M1型丙酮酸激酶）和PKM2（M2型丙酮酸激酶）的表达变化；最后，利用葡萄糖和乳酸检测试剂盒分析过表达和敲低RBMX对膀胱癌细胞糖酵解的影响。结果 RT-qPCR和Western blotting结果显示，过表达RBMX膀胱癌细胞的mRNA和蛋白表达水平显著高于阴性对照组（P<0.05），敲低RBMX膀胱癌细胞的mRNA和蛋白表达水平显著低于阴性对照组（P<0.05）。过表达RBMX明显抑制膀胱癌细胞的增殖、克隆形成、迁移和侵袭，敲低RBMX则作用相反。Western blotting实验结果显示，过表达RBMX使PKM1表达上升，PKM2表达下降，敲低RBMX则作用相反。葡萄糖消耗及乳酸生成实验表明，过表达RBMX均能抑制膀胱癌细胞葡萄糖消耗及乳酸生成（P<0.05），敲低RBMX均能促进膀胱癌细胞葡萄糖消耗及乳酸生成（P<0.05）。结论 RBMX通过下调PKM2抑制膀胱癌的发生发展和糖酵解能力，有望成为膀胱癌诊断和治疗的潜在分子靶标。

关键词: X连锁RNA结合基序蛋白；M2型丙酮酸激酶；膀胱癌；PKM2；糖酵解

Abstract: Objective To investigate the role of RNA-binding motif protein X- linked (RBMX) in regulating the proliferation, migration, invasion and glycolysis in human bladder cancer cells. Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown, respectively, and successful cell modeling was verified using RT-qPCR and Western blotting. Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony- forming assay, and cell migration and invasion abilities were determined using Transwell experiment. The expressions of glycolysis- related proteins M1 pyruvate kinase (PKM1) and M2 pyruvate kinase (PKM2) were detected using Western blotting. The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits. Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown. RBMX overexpression significantly inhibited the proliferation, clone formation, migration and invasion of bladder cancer cells, while RBMX knockdown produced the opposite effects. Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2, while RBMX knockdown produced the opposite effects. Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression (P<0.05) but increased significantly following RBMX knockdown (P<0.05). Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression, suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Key words: RNA-binding motif protein X-linked; M2 pyruvate kinase; bladder cancer; PKM2; glycolysis