miRNA-138与EZH2在肾透明细胞癌发生发展中作用机制的研究

目的:探讨miRNA-138及其调控靶基因EZH2在肾透明细胞癌(RCC)发生发展中的相关作用机制。方法:利用第二代大规模平行测序技术对10例RCC及对应癌旁组织进行全基因组测序,在肾癌组织中筛选出表达显著上调的miRNA-138;其次在人肾癌细胞株786-O、ACHN、Caki-1、769-P、os-rc-2及对照细胞人胚肾细胞293T中对miRNA-138进行转染,采用划痕实验检测miRNA-138对细胞株侵袭和迁移能力的影响;进而用多种预测软件预测靶基因,对预测的候选靶基因EZH2进行PCR验证。结果:发现miRNA-138在RCC中表达上调,并能够显著促进人肾癌细胞株786-O、ACHN的迁移;而其候选靶基因EZH2的表达在临床组织标本中显著上调;相应蛋白变化需要进一步验证。结论:EZH2表达在RCC中明显高于正常组织,提示EZH2可能是RCC一个敏感的生物学标志。推测miRNA-138与其靶基因EZH2可能参与肿瘤代谢和细胞凋亡等信号通路,在RCC的发生发展中发挥重要作用。

Study on mechanism of miRNA-138 and EZH2 in the occurrence and development of renal clear cell carcinoma

Objective：To investigate the mechanism of miRNA-138 and its target gene enhancer of zeste homolog 2 （EZH2） in the occurrence and development of renal clear cell carcinoma （RCC）. Method：The second-generation massively parallel signature sequencing examining the miRNA expression in tumor and the corresponding paracancerous tissues of ten RCC patients was used to screen that miRNA-138 was notably up-regulated in RCC tissues. The scratch assay was performed to detect the invasion and migration, after that the renal cell carcinoma cell lines, 786-0, ACHN, Caki-1, 769-P, os-rc-2 and the control cell line 293T were transfected with miRNA-138. Finally bioinformatics analysis was used to predict EZH2 as a candidate target gene and PCR was performed to val idate it. Result：MiRNA-138 was significantly up-regulated in RCC and could promote the migration of 786-0 and ACHN cell lines remarkably. While its candidate target gene EZH2 was dramatically up-regulated in RCC speci- mens and its corresponding protein change need further validation. Conclusion：The expression of EZH2 is obviously higher in RCC than in the adjacent tissue, which indicates that EZH2 may be a sensitive biomarker in RCC. MiRNA-138 and its target gene EZH2 may be involved in the tumor metabolism and apoptosis signaling pathways and play important roles in the occurrence and development of RCC.