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作 者:郝美君[1,2] 郑素军[2] 丁惠国[2] 黄爱龙[1]
机构地区:[1]重庆医科大学,重庆400016 [2]首都医科大学附属北京佑安医院,北京100069
出 处:《生物医学工程学杂志》2011年第4期784-789,803,共7页Journal of Biomedical Engineering
基 金:973计划前期研究专项基金资助项目(2007CB516810);国家自然科学基金资助项目(30800979);北京市科技新星课题资助项目(2007B055);北京自然科学基金资助项目(7102085)
摘 要:为了寻找与HBV感染相关的microRNA并初步探讨其作用机制,首先,我们前期研究发现在转染HBV表达质粒的HepG2细胞内,miR-122上调表达,推测miR-122与HBV的感染有关;在此基础上,本研究进一步将miR-122和表达HBV的质粒pCH9-HBV1.1共转染HepG2细胞,Southern blot检测发现miR-122可抑制HBV的复制。利用计算机软件miRanda预测表明,HBx可能是miR-122的作用靶序列;进一步利用荧光素酶报告基因系统和Western blot检测靶基因HBx蛋白表达改变,验证miR-122对HBx表达的调节作用。最终推测miR-122可能通过调节HBx的表达而影响HBV的复制。In order to find microRNA associated with HBV infection and to explore the mechanism of the infection,first of all,we found in our preliminary study that in HepG2 cells transfected with HBV expression plasmid,miR-122 expression was up-regulated,suggesting that miR-122 was related to the HBV infection.On this basis,in the present study,miR-122 and pCH9-HBV1.1 plasmid were cotransfected into HepG2 cells.Southern blot detection result showed that miR-122 can inhibit HBV replication.Using MiRanda computer software,HBx was predicted to be the target sequence of miR-122;Luciferase reporter gene system and Western blot detection of HBx protein expression changes were further used to verify the HBx expression regulated by miR-122.And finally,it can be speculated that miR-122 may affected HBV replication by regulating the expression of HBx.
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