核糖核苷酸还原酶调节亚基M2对膀胱癌细胞增殖和侵袭的影响

Effect of RRM2 on proliferation and invasion of bladder cancer cells

目的探讨核糖核苷酸还原酶调节亚基M2(ribonucleotide reductase regulatory subunit M2,RRM2)在膀胱癌细胞中的生物学功能及其机制。方法采用TCGA及Oncomine数据库分析膀胱癌患者RRM2的表达,生存分析采用K-M Plot在线分析网站。收集2019年6月至2020年12月在本院泌尿外科手术患者的膀胱癌组织和癌旁组织,通过qRT-PCR检测其RRM2的表达情况。采用膀胱癌细胞系(5637、T24、Ej、BIU-87)进行细胞实验;选取高表达RRM2的细胞进行沉默实验,采用CCK-8、Transwell检测RRM2对膀胱癌细胞增殖、侵袭的影响,Western blot检测wnt/β-catenin信号通路相关蛋白的表达。结果 RRM2在膀胱癌患者中表达升高(差异表达倍数Log2FC=4.546,P<0.01),高表达的RRM2与患者不良预后及较差的总体生存率相关。体外细胞实验显示,与5637和T24细胞相比,siRNA转染沉默RRM2后72、96 h显著抑制膀胱癌细胞的增殖(P<0.01);并显著减弱膀胱癌细胞的侵袭能力(P<0.01)。沉默RRM2后,总β-catenin表达水平显著降低,Wnt/β-catenin信号通路的下游蛋白,包括在肿瘤进展中起重要作用的细胞周期蛋白D1和c-Myc也显著降低。结论 RRM2可促进膀胱癌细胞的增殖和侵袭,可能靶向wnt/β-catenin通路发挥促癌作用。

Objective To explore the biological function and mechanism of ribonucleotide reductase regulatory subunit M2(RRM2) in bladder cancer cells. Methods The Cancer Genome Atlas(TCGA) and Oncomine databases were used to analyze the expression of RRM2 in patients with bladder cancer, and the K-M Plot online analysis was used for survival analysis. The bladder cancer tissues and adjacent tissues from the patients undergoing urological surgery in our hospital from June 2019 to December 2020 were collected and detected for the expression of RRM2 by qRT-PCR. CCK-8 and Transwell assays were employed to evaluate the effects of RRM2 on proliferation and invasion in bladder cancer cell lines(5637, T24, Ej and BIU-87 cells). Then the effect of RRM2 silence was determined with CCK-8 and Transwell assays in the cells with higher expression of RRM2, and the expression levels of Wnt/β-catenin signaling pathway related proteins were measured with Western blotting. Results The expression of RRM2 was increased in patients with bladder cancer(differential expression multiple Log2FC=4.546, P<0.01), and the patients with high expression of RRM2 were associated with poor prognosis and poor overall survival. In vitro cell experiments showed that RRM2 silence significantly inhibited the proliferation and reduced the invasion of bladder cancer RRM2 cells in 72 and 96 h after siRNA transfection when compared with the 5637 and T24 cells(P<0.01). The silence also significantly decreased the expression levels of total β-catenin and Wnt/β-catenin downstream proteins cyclin D1 and c-myc, which played important roles in tumor progression. Conclusion RRM2 can promote the proliferation and invasion of bladder cancer cells, and may target the Wnt/β-catenin pathway to promote bladder cancer.