microRNA-204在鼻咽癌中的表达及生物学意义研究

Expression and biological significance of microRNA-204 in nasopharyngeal carcinoma

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q RT-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q RT-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 Micro RNA-204 in nasopharyngeal carcinoma（ NPC）. Methods： q RT-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 m RNA 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.