中国病理生理杂志2016，Vol.32Issue(11)：1958-1965,8.DOI:10.3969/j.issn.1000-4718.2016.11.007
HIF-1α介导了间歇低氧对人肺腺癌A549细胞体外生物学行为的影响∗
HIF-1αmediates effect of intermittent hypoxia on biological behavior of A549 cells in vitro
摘要
Abstract
[ ABSTRACT] AIM:To investigate whether hypoxia-inducible factor 1α( HIF-1α) mediates the effect of intermit-tent hypoxia on A549 cell viability, apoptosis and invasive ability in vitro. METHODS: A549 cells were transfected with HIF-1α-siRNA and cultured under intermittent hypoxia. The expression of HIF-1αand its downstream genes, such as Bcl-2, Bax, P53, P21 and VEGF at mRNA and protein levels was determined by real-time PCR and Western blot. The viabili-ty of the A549 cells was measured by MTT assay. The apoptosis and cell cycle distribution of the A549 cells were examined by flow cytometry. The invasive ability of the A549 cells was detected by transwell test. RESULTS:The expression levels of HIF-1α, Bcl-2 and VEGF in non-HIF-1α-siRNA transfected A549 cells cultured in intermittent hypoxia environment [ blank control group ( IHC ) , empty vector control group ( IHE ) and negative control group ( IHN ) ] were higher than those in the A549 cells in normoxia group ( RA) , but the expression levels of Bax and P21 were lower than those in RA group (P<0. 05). The siRNA-mediated HIF-1α gene silencing [intermittent hypoxia silenced group (IHS)] resulted in obvious down-regulation of HIF-1α, Bcl-2 and VEGF, and significant increase in the protein expression of P21 and Bax (P<0. 05). The expression level of P53 in intermittent hypoxia groups was significantly higher than that in RA group, and no significant difference of P53 expression in different intermittent hypoxia groups was observed. Compared with normoxia, intermittent hypoxia resulted in significantly enhanced cell viability, decreased apoptosis, and enhanced invasive ability of non-HIF-1α-siRNA transfected A549 cells (P<0. 05). The siRNA-mediated HIF-1αgene silencing resulted in significant cell viability inhibition, elevated apoptotic rate and decreased invasive ability under hypoxic condition (P<0. 05). CON-CLUSION:Intermittent hypoxia promotes the viability and invasion of A549 cells by HIF-1α-mediated downstream gene expression. HIF-1α gene silencing inhibits A549 cell growth and invasion under intermittent hypoxia by inhibition of HIF-1α signal pathways in vitro.关键词
间歇低氧/缺氧诱导因子1α/RNA干扰/A549细胞Key words
Intermittent hypoxia/Hypoxia-inducible factor 1α/RNA interference/A549 cells分类
医药卫生引用本文复制引用
杨士芳,李晓明,安弘,高兴林..HIF-1α介导了间歇低氧对人肺腺癌A549细胞体外生物学行为的影响∗[J].中国病理生理杂志,2016,32(11):1958-1965,8.基金项目
国家自然科学基金资助项目(No.81300034) （No.81300034）
广东省科技计划项目(No.2014A020212677) （No.2014A020212677）
广东省医学科研基金项目(No. A2015029) （No. A2015029）
