microRNA-204在鼻咽癌中的表达及生物学意义研究

摘要

目的：：研究微小RNA 204(microRNA-204,miR-204)在鼻咽癌(Nasopharyngeal carcinoma,NPC)中的表达情况及生物学意义。方法：收集2012年3月至2014年3月间于我院耳鼻喉头颈外科行组织病理活检的NPC组织及对应癌旁鼻黏膜组织共43例,运用qRT-PCR技术检测miR-204在NPC组织及癌旁组织中的表达水平,统计分析miR-204表达水平与患者临床病理资料间的相关性；免疫组化检测鼻咽癌组织中miR-204下游潜在靶点Bcl-2及SIRT1蛋白的表达水平,统计分析Bcl-2及SIRT1蛋白与miR-204表达间的相关性；采用人工合成的miR-204模拟物( miR-204 mimics)转染人鼻咽癌CNE-2细胞,分别采用qRT-PCR及Western blot检测转染后Bcl-2及SIRT1 mRNA和蛋白的表达变化。结果：miR-204在NPC组织中表达水平显著低于对应癌旁鼻黏膜组织(P<0．05)；miR-204低表达与肿瘤淋巴结转移(P<0．05)及高TNM分期(Ⅲ+Ⅳ期,P<0．05)显著相关；在鼻咽癌组织中miR-204与其下游潜在靶点Bcl-2及SIRT1的表达呈显著负相关关系(P<0．05),miR-204转染人鼻咽癌CNE-2细胞后可显著降低细胞内Bcl-2及SIRT1 mRNA和蛋白的表达水平(P<0．05)。结论：miR-204在鼻咽癌组织中表达下调并与肿瘤恶性临床病理特征有关,miR-204可能通过下调Bcl-2及SIRT1的表达来抑制鼻咽癌的发生、发展过程。%Objective:To investigate the expression and biological significance of MicroRNA-204 in nasopharyngeal carcinoma (NPC). Methods: qRT-PCR was applied to detect the relative expression of miR-204 in 43 paired nasopharyngeal carcinoma in comparison to the normal nasal mucosa tissues. Pearson chi-square test was used to analyze the relationship between the miR-204 expression and clinical features. The expressions of Bcl-2 and SIRT1 were measured by immunohistochemistry ( IHC ) , Spearman correlation analysis was used to analyze the relationship between miR-204 and Bcl-2,as well as SIRT1. We then transfected the miR-204 mimics into CNE-2 cells,then the Western blot was used to detect the expression of Bcl-2 and SIRT1,which were considered as the potential targets of miR-204. Results:The relative expressions of miR-204 was significantly downregulated in NPC tissues compared to those of the matched normal tumor-adjacent tissues(P<0. 05). Low expression of miR-204 was significantly associated with lymphatic metastasis(P<0. 05) and advanced TNM stage(Ⅲ+Ⅳ,P<0. 05). The expressions of Bcl-2 and Sirt1 in lower miR-204 level group were both higher than in higher miR-204 level group ( P<0. 05 ) . Both the mRNA and protin expression in CNE-2 cells were down-regulated after transfection. Conclusion: Low expression of miR-204 is related to the malignant clinicopathological features in NPC tissues,and miR-204 may through down-regulate Bcl-2 and SIRT1 to suppress NPC genesis and development.