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Effects of RNAi on hypoxia inducible factor-1α activity and proliferation of hypoxic pulmonary artery smooth muscle cells in rat
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- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 1
- DOI:
- 10.3321/j.issn:0371-0874.2006.01.012
- Key Word:
- 肺动脉;平滑肌细胞;低氧诱导因子-1α;RNA干扰;细胞增殖
Abstract： 本研究应用基因克隆技术,将合成的发卡样特异性低氧诱导因子-1α(hypoxiainduciblefactor-1 alpha,HIF-1α)干扰寡核苷酸(siRNA)序列插入真核表达载体中,构建出特异性HIF-1αt基因RNA干扰(RNAi)真核表达载体.采用组织块种植法,原代培养大鼠肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs),将构建出的特异性HIF-1α RNAi真核表达载体转染到PASMCs;分别在常氧和低氧下进行细胞培养,采用RT-PCR检测PASMCs HIF-1α mRNA表达水平,用MTT和流式细胞仪检测细胞增殖水平,探讨低氧条件下HIF-1α RNAi真核表达载体对PASMCs增殖的影响.结果表明,低氧培养48 h后,正常PASMCs和转染了HIF-1α siRNA阴性表达载体的细胞增殖显著,HIF-1αmRNA表达水平也显著升高;而转染了HIF-1α siRNA阳性表达质粒的细胞增殖不显著,HIF-1αmRNA表达水平较低.结果提示:HIF-1αRNAi真核表达载体能显著干扰培养的PASMCs HIF-1α mRNA表达,同时抑制低氧环境下PASMCs的增殖.
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