摘要
目的:探讨miRNA-93在膀胱癌中的表达及对人膀胱癌细胞株T24细胞生物学特性的影响及其作用机制。方法:选取郑州大学附属南阳市中心医院泌尿外科2010年5月至2016年5月收治的79例膀胱癌患者病理组织标本及配对癌旁正常组织,通过qRT-PCR检测miRNA-93的表达水平。沉默miRNA-93后,采用CCK-8法、Transwell实验和流式细胞术分别检测细胞增殖和迁移能力的变化及细胞的凋亡情况;Western blotting检测AKT/p-AKT、GSK3β/p-GSK3β蛋白表达水平的变化。结果:miRNA-93在膀胱癌组织及癌细胞中高表达,且表达水平与癌灶大小、淋巴结转移、病理分级及T分期有关(均P<0.05)。沉默miRNA-93后,T24细胞的增殖能力显著降低(P<0.05),沉默miRNA-93促进T24细胞的凋亡并抑制其迁移;同时,沉默miRNA-93后p-AKT和p-GSK3β的蛋白表达水平均显著下降(P<0.05)。结论:miRNA-93能够促进人膀胱癌细胞株T24细胞的增殖和迁移,并抑制凋亡,其作用机制可能与下调p-AKT、p-GSK3β蛋白表达有关,提示miRNA-93可以作为诊断和靶向治疗膀胱癌的潜在作用位点。
Objective：To investigate the expression of miRNA-93 and its effect on the cellular biological characteristics of bladder cancer cell line T24, and to explore the underlying mechanism.Methods：The cancer tissues and paired adjacent normal tissues from 79 patients with bladder cancer treated in Nanyang Center Hospital Affiliated to Zhengzhou University between May 2010 and May 2016 were selected for this study. The expressions of miRNA-93 were detected by Real-time PCR. After silencing of miRNA-93, the cell proliferation, apoptosis and migration were measured by CCK-8 assay, Flow cytometry and Transwell assay, respectively. The protein expressions of AKT/p-AKT and GSK3β/p-GSK3β were measured by Western blotting assay. Results： MiRNA-93 was highly expressed in both bladder cancer tissues and cancer cells （P〈0.05）, and its expression level was closely related to tumor size, lymph node metastasis, pathological grade stage and TNM classification （P〈0.05）. CCK-8 assay and cell cycle analysis showed that knockdown of miRNA-93 significantly suppressed T24 cell proliferation （P〈0.05）, and Flow Cytometry AnnexinV / PI double staining showed miRNA-93 silencing promoted T24 cell apoptosis （P〈0.05）. And Transwell assay showed that knockdown of miRNA-93 suppressed T24 cell migration. Furthermore, Western blotting showed that the protein expressions of p-AKT and p-GSK3β were significantly decreased after down-regulation of miRNA-93 （P〈0.05）. Conclusion：miRNA-93 could enhance cell proliferation and migration, and suppress cell apoptosis of human bladder cancer T24 cell line. And its mechanism might be related to the down-regulation of p-AKT and p-GSK3β protein expression, suggesting that miRNA-93 could be used as a potential new target for diagnostic and targeted therapy of bladder cancer.
作者
张红
刘念
李征
乔保平
ZHANG Hong LIU Nian LI Zheng QIAO Baoping(Department of Nephropathy Rheumatology Second Department of Orthopaedics Department of Urology, Nanyang Center Hospital Affiliated to Zhengzhou University, Nanyang 473000, Henan, China Department of Urology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2017年第2期139-144,共6页
Chinese Journal of Cancer Biotherapy
基金
河南省基础与前沿技术研究计划基金资助(No.142300410034)~~
