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论文摘要

食管鳞癌中miRNA224调控RKIP基因表达及miRNA224基因启动子区甲基化的研究

STMiRNA224 Regulation Role on RKIP Gene Expression and Methylation of miRNA224 Gene Promoter Region inEsophageal Squamous Cell Carcinoma

作者:牛中喜, 杨晓龙, 刘明佳等

Author:NIU Zhong-xi, YANG Xiao-long, LIU Ming-jia. et al

收稿日期:          年卷(期)页码:2016,47(4):468-472

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:miRNA224 RKIP 食管鳞癌 甲基化

Key words:miRNA224 RKIP Esophageal squamous cell carcinoma Methylation

基金项目:

中文摘要

目的 研究食管鳞癌组织中Raf激酶抑制蛋白(RKIP)、miRNA224表达情况,miRNA224 对RKIP的靶向作用,以及miRNA224基因上游启动子区甲基化水平。方法 在食管鳞癌组织及癌旁对照组织临床标本中,用免疫组化法检测RKIP蛋白的表达;用荧光定量PCR法检测miRNA224的表达,荧光素酶报告基因检测miRNA224对RKIP的靶向作用;亚硫酸氰盐测序PCR(BSP)法检测食管鳞癌及癌旁对照组织miRNA224基因上游启动子区甲基化水平。结果 在食管鳞癌组织中,RKIP低表达,miRNA224高表达;在癌旁对照组织中,RKIP高表达,miRNA224低表达。荧光素酶报告基因验证miRNA224可靶向抑制RKIP 3′UTR表达。BSP法显示食管鳞癌组织miRNA224上游启动子区呈低甲基化状态,癌旁对照组织呈高甲基化状态。结论 食管鳞癌组织RKIP表达下降、miRNA224基因启动子甲基化状态降低,miRNA224表达上升,RKIP是miRNA224下游作用的靶点,可能同食管鳞癌发生密切相关。

英文摘要

Objective To study the expression levels of tumor suppressor gene RIKP and miRNA224 in esophageal squamous cell carcinoma (ESCC) tissues. To determine whether miRNA224 targets RKIP and the methylation status of miRNA224 gene promoter region in esophageal carcinoma. Methods The expression levels of RKIP and miRNA224 in ESCC and normal tissue were detected by using immunohistochemistry and real-time qPCR, respectively. Luciferase assay was used to determine the targeting of miRNA224 to RKIP. The methylation status of miRNA224 promoter region was studied by bisulfite sequencing PCR (BSP). Results In 40 cases of ESCC, RKIP expression was significantly lower than that of normal tissue; miRNA224 expression was higher in ESCC than in paracancerous tissue. Luciferase assay showed that miRNA224 targets RKIP 3′UTR thus inhibit its expression. The miRNA224 gene promoter region was hypomethylated in ESCC. Conclusion Compared with normal tissue, in ESCC, RKIP was downregulated, while miRNA224 was upregulated, and the promoter region of miRNA224 gene was hypomethylated. RKIP is the target of miRNA224, which may be closely related to esophageal squamous cell carcinoma.

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