解放军医学杂志2017，Vol.42Issue(4)：320-325,6.DOI:10.11855/j.issn.0577-7402.2017.04.09
siRNA沉默EPAS1基因对低氧条件下大鼠PASMCs增殖的影响
Effects of siRNA on EPAS1 activity and proliferation of hypoxic pulmonary artery smooth muscle cells in rats
摘要
Abstract
Objective This study explored whether EPAS1 gene is involved in the proliferation of pulmonary arterial smooth muscle cells (PASMCs) during hypoxia when EPAS1 gene expression was interfered by small interfering RNA (siRNA).Methods The rat primary pulmonary artery smooth muscle cells were cultured and identified by immunofluorescence.The specific lipidosomes of EPAS1 siRNA were constructed and transfected into PASMCs,and the best targets were selected from the three interfering targets.The transfected PASMCs were cultured in hypoxia (37℃,5％ CO2,2％ O2) or normoxia (37℃,5％ CO2,20％ O2) for 24h,48h and 72h,respectively.The PASMCs proliferation was detected by CCK-8 assay.The protein expression of EPAS 1 and vascular endothelial growth factor (VEGF) were determined by Western blotting to investigate the effect of silencing EPAS1 gene expression on the proliferation of PASMCs under hypoxic condition.Results The specific liposomes ofEPAS1 siRNA were successfully constructed and transfected into PASMCs,and the best interfering target were selected from the three interference targets.The proliferation of PASMCs was increased and the protein expression of VEGF was up-regulated in the PASMCs under hypoxic condition.Under hypoxic or normoxic condition,PASMCs proliferation and VEGF protein expression of PASMCs were suppressed by EPAS 1 siRNA.Conclusion EPAS 1 gene might be involved in the proliferation of rat PASMCs by regulating VEGF protein level under hypoxic condition.关键词
EPAS1基因沉默/低氧/肺动脉平滑肌细胞/增殖Key words
EPAS1 gene/hypoxia/pulmonary arterial smooth muscle cells/proliferation分类
医药卫生引用本文复制引用
孙莉,杨全余,嘎琴,靳国恩..siRNA沉默EPAS1基因对低氧条件下大鼠PASMCs增殖的影响[J].解放军医学杂志,2017,42(4):320-325,6.基金项目
国家自然科学基金(81160243) （81160243）
青海省留学回国人员科技活动项目(青人社厅函[2011]518号)This work was supported by the National Natural Science Foundation of China (8160243),and the Scientific and Technological Activities Project of Chinese Overseas Returnees (2011-518) （青人社厅函[2011]518号）
