| 秦国伟 毛文君 王谦 陈静瑜 聂晓伟.MicroRNA-18a通过HIF-1-alpha调控缺氧引起的人肺动脉平滑肌细胞增殖[J].,2016,16(16):3023-3027 |
| MicroRNA-18a通过HIF-1-alpha调控缺氧引起的人肺动脉平滑肌细胞增殖 |
| MicroRNA-18a Regulates the Proliferation Pulmonary Artery Smooth MuscleCells Through HIF-1-alpha |
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| DOI： |
| 中文关键词: 肺动脉平滑肌细胞 MicroRNA-18a 缺氧 缺氧诱导因子-1alpha |
| 英文关键词: Human pulmonary artery smooth muscle cells Hypoxia microRNA-18a HIF-1alpha |
| 基金项目:国家自然科学基金项目(81500039)；南京医科大学科技发展基金项目(2015NJMU148) |
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| 中文摘要: |
| 目的：研究microRNA-18a (miR-18a) 对缺氧引起的人肺动脉平滑肌细胞(human pulmonary artery smooth muscle cells，
hPASMCs)增殖的调控作用及其可能机制。方法：体外培养hPASMCs，分为未转染组、miR-18a 模拟物对照组、miR-18a 模拟物组、
miR-18a 抑制剂对照组、miR-18a 抑制剂组、siRNAcontrol组、siHIF-1-alpha组、miR-18a 抑制剂和siHIF-1-alpha共转染组。分别于常氧(21%
O2)和低氧(3%O2)作用24小时。采用CCK-8 法检测细胞的增殖情况，萤光素酶报告基因系统验证缺氧诱导因子-1-alpha(HIF-1alpha)是
否为miR-18a 的靶基因，并通过western-blot 以及实时荧光定量PCR 技术检测相关蛋白和基因的表达。结果：缺氧可促进
hPASMCs 增殖，使miR-18a 表达减少；miR-18a 模拟物可抑制hPASMCs 增殖，而miR-18a 抑制剂可促进hPASMCs 增殖；抑制
miR-18a 可使HIF-1-alpha的表达上调。同时抑制miR-18a 和HIF-1-alpha，可使miR-18a 对hPASMCs 增殖调控的能力消失。结论：缺氧通
过抑制miR-18a，上调HIF-1-alpha的表达，促进hPASMCs增殖。 |
| 英文摘要: |
| Objective:To study the effects of microRNA-18a (miR-18a) on human pulmonary artery smooth muscle cells
(hPASMCs) proliferation and the mechanisms involved.Methods:hPASMCs were divided into eight groups, including: non-transfected
group, miR-18a mimic control group, miR-18a mimic group, miR-18a inhibitor control group, miR-18a inhibitor group, siRNA control
group, siHIF-1-alpgha group, miR-18a inhibitor and siHIF-1-alpha co-transfected group. Cells were subjected to normoxia (21%O2) and hypoxia
(3%O2) for 24 h. Cell viability (CCK-8), miRNA-mRNA interactions (HIF-1-alpha3' untranslated region Luciferase reporter assays), gene
expression (quantitative PCR and western-blot) were assessed under normoxic or hypoxic conditions.Results:Hypoxia promoted
hPASMCs proliferation but inhibited miR-18a expression. Ectopic expression of miR-18a in hPASMCs reduced proliferation, whereas
miR-18a inhibition in hPASMCs promoted proliferation. We identified HIF-1-alpha as a direct target of miR-18a. Modulating miR-18a
expression significantly affected HIF-1-alpha expression. The capacity of miR-18a on hPASMCs proliferation was attenuated by inhibiting
miR-18a and HIF-1-alphatogether.Conclusion:Together, these data support a role of miR-18a in hypoxia-induced hPASMCs proliferation
through a HIF-1-alpha -dependent pathway. |
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