摘要
目的研究miR-214在肺动脉高压发病机制中的调控作用。方法应用逆转录聚合酶链式反应(qRT-PCR)方法检测miR-214在肺动脉高压患者血清及缺氧诱导肺动脉平滑肌细胞中表达;通过EdU渗入法以及划痕实验研究miR-214对肺动脉平滑肌细胞增殖及迁移的影响;对miR-214靶基因预测并验证。结果肺动脉高压患者血清中miR-214显著高于正常人血清(p<0.001);经过缺氧处理肺动脉平滑肌细胞明显促进miR-214表达(p<0.01);miR-214促进缺氧条件下的肺动脉平滑肌细胞增殖、迁移;双荧光素酶报告系统证明miR-214能够与缺氧诱导因子1α抑制因子(HIF1AN)的3'-UTR结合;HIF1AN为miR-214在肺动脉平滑肌细胞中的靶向基因。结论miR-214可能通过靶基因HIF1AN参与肺动脉高压调节机制。
Objective To explore the function of miR-214 and potential molecular mechanisms in pulmonary arterial hypertension. Methods miR-214 expression was identified by qRT-PCR in serum of patients with PAH associated with congenital heart disease, and the pulmonary artery smooth muscle cells （PASMC） exposed to hypoxia. Then assessed the role of miR-214 in modulating cell proliferation and migration in hypoxia-induced PASMC by EdU incorporation and wound assay. The target genes of miR-214 were predicted by targetscan soft and validated. Results miR-214 was significantly increased in the serum of CHD-PAH patient as compared with controls as well as in PASMC exposed to hypoxia （p〈0.001）. Through functional analysis displayed that miR-214 significantly enhanced hypoxia-induced PASMC proliferation and migration. HIFIAN was identified as a direct target of miR-214 by 3＇-UTR luciferase assay and further confirmed at protein levels. Conclusions The study demonstrates that miR-214 was involved in regulating PAH by targeting HIF1AN.
作者
张洪亮
贾阿娜
陈宇
王慧
刘春辉
刘擎
彭鹏
张峰
贺兆发
ZHANG Hong-liang JIA A-na CHEN Yu WANG Hui LIU Chun-hui LIU Qing PENG Peng ZHANG Feng HE Zhao-fa(Department of Cardiology the First Affiliated Haspital of Jiamusi University, Jiamusi, Heilongjiang, 154002, China.)
出处
《中国分子心脏病学杂志》
CAS
2016年第5期1828-1831,共4页
Molecular Cardiology of China
基金
黑龙江省自然科学基金项目(H201364)
黑龙江省卫生厅项(2011-355)
