微小RNA-373在肝门部胆管癌细胞中的达和靶向甲基CpG结合蛋白2的作用  

Expression and targeting to methyl-CpG-binding domain 2 of microRNA-373 in hilar cholangiocarcinoma

作　　者：陈勇军[1] 田礼[1] 胡走肖[1] 刘永[1] 邹声泉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院胆胰外科,武汉430030

出　　处：《中华实验外科杂志》2013年第11期2255-2257,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81071998）;湖北省自然科学基金杰出青年基金资助项目（2012FFA012）

摘　　要：目的 分析微小RNA（miR）-373在肝门部胆管癌的表达与临床病理因子之间的关系,探讨miR-373靶向甲基CpG结合蛋白2（MBD2）-3’端非翻译区（3＇ UTR）的机制.方法 TaqMan miRNA Assay检测miR-373的表达,采用QuantiTect_Primer Assays和Western blot法分析甲基CpG结合蛋白（MBPs） mRNA和蛋白水平；双荧光素酶报告基因检测miR-373对MBD2-3＇ UTR的靶向作用.结果 miR-373在肝门部胆管癌中上调2.94倍（P＜0.01）.与高表达组比较,miR-373低表达与肿瘤细胞低分化（P＜0.05）、高分期（P＜0.05）、短总生存和无病生存时间（P＜0.05）相关.miR-373低表达肿瘤组织中,MBD2的mRNA和蛋白表达分别上调3.75倍（P＜0.01）、2.34倍（P＜0.05）；miR-373前体抑制MBD2-3＇ UTR荧光素酶活性1.79倍；外源性miR-373和anti-miR-373分别下调QBC939细胞、上调人肝内胆管上皮细胞（HIBEpic）中MBD2表达2.56倍（P＜0.01）和2.13倍（P＜0.01）.结论 miR-373通过结合在MBD2-3＇ UTR,抑制MBD2表达.肝门部胆管癌中,miR-373下调导致MBD2表达增高,与肿瘤进展和预后密切相关.Objective To study the expression and targeting to methyl-CpG-binding domain 2 （MBD2） of microRNA （miR） -373 in hilar eholangiocarcinoma. Methods The miR-373 expression was detected by using TaqMan miRNA assay. The mRNA and protein expression of methyl-CpG binding domain proteins （MBPs） was examined by using QuantiTect_Primer assays and Western blotting, respectively. The targeting at MBD2-3 ＇ UTR by miR-373 was evaluated by using dual-luciferase reporter gene assay. Re- suits The miR-373 expression was decreased 2. 94 times and was closely associated with poor cell differ- entiation （ P 〈 0. 05 ）, advanced clinical stage （ P 〈 0. 05 ）, and shorter survival （ P 〈 0. 05 ） in hilar eholangioearcinoma. In high miR-373 group, the MBD2 mRNA and protein expression was increased 3.75 times （ P 〈 0.01 ） and 2. 34 times （ P 〈 0.05 ）, respectively. Precursor miR-373 inhibited the luciferase activity of MBD2-3 ＇ UTR by 1.79 times. Exogenous miR-373 and anti-miR-373 suppressed MBD2 in QBC939 cells by 2. 56 times （ P 〈 0. 01 ）, and enhanced MBD2 in human intrahepatic biliary epithelial cells（HIBEpic） by 2. 13 times （ P 〈 0. 01 ）. Conclusion miR-373 is one negative regulator of MBD2. The increase of MBD2 induced by down-expression of miR-373 plays an important role in development and prognosis of hilar eholangiocarcinoma.

关 键 词：微小RNA-373 甲基化CpG结合蛋白2 肝门部胆管癌 

分 类 号：R735.8[医药卫生—肿瘤]