miR-124在肾癌中的表达以及对肾癌786-O细胞生物学行为的影响

Expression of miR-124 and its effect on biological function in renal cell carcinoma 786-O cells

目的探讨肾癌中miR-124的表达及其与临床病理特征的关系;观察肾癌786-O细胞中miR-124对细胞增殖、迁移和侵袭的作用,以及对EZH2表达的影响。方法采用RT-PCR法检测62例肾癌组织及相应癌旁正常组织临床样本中miR-124的表达水平,并分析miR-124的表达水平与肾癌患者临床病理特征之间的相关性;采用MTT法和Transwell实验观察细胞增殖、迁移和侵袭水平的改变;使用在线数据库及荧光素酶报告实验预测并验证miR-124的靶基因,并采用Western blot法检测细胞中EZH2蛋白的表达水平。结果与癌旁正常组织相比,miR-124在肾癌组织中表达明显降低,其表达水平与肿瘤的组织学分级、临床分期以及淋巴结转移密切相关;过表达miR-124可显著减少肾癌786-O细胞增殖、迁移和侵袭的水平;在线数据库及荧光素酶报告实验预测并验证了EZH2是miR-124的下游靶基因,过表达miR-124可使EZH2蛋白水平表达降低。结论 miR-124在肾癌中低表达,其可能通过靶向EZH2的表达,抑制肾癌786-O细胞增殖、迁移和侵袭的能力。

Objective To investigate the expression of miR-124 and its relationship with clinicopathological features in kidney cancer tissues, and to observe the roles of miR-124 on the proliferation,migration an invasion in renal cell carcinoma 786-O cells and the influence on EZH2.Methods RT-PCR method was used to detect the expression level of miR-124 in 62 cases of kidney cancer tissues and adjacent normal tissues,and the correlation between the expression of miR-124 and the clinicopathological characteristics was analyzed. MTT and Transwell experiments were evaluated to observe the effects of miR-124 in cell proliferation, migration and invasion. The target gene of miR-124 was predicted and validated by online database and luciferase Report experiments, and the protein expression level of EZH2 were determined by western blot(WB) method. Results Compared with normal tissues, the expression of miR-124 in kidney cancer tissues decreased significantly,and its expression level was closely related to histological grade, clinical stage and lymph node metastasis of the tumor. Overexpression of miR-124 could significantly reduce the proliferation, migration and invasion of 786-O cells in renal cell carcinoma. Online database and luciferase reporter experiments predicted and verified that EZH2 is the downstream target gene of miR-124. Overexpression of miR-124 can reduce the protein expression level of EZH2. Conclusion MiR-124 has a lower expression level in kidney cancer tissues, and it may play an important role in reducing the level of cell proliferation, migration and invasion of 786-O cells by targeting EZH2.