缺氧对肺癌细胞系A-549表达缺氧诱导因子1α和2α的影响
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摘要
目的探讨常氧状态和不同缺氧持续时间、不同氧浓度下肺癌A-549细胞中缺氧诱导因子(HIF-1α、HIF-2α)表达水平的变化规律。方法将A549细胞分为常氧组、时间控制组、氧浓度控制组,采用Western blot分别检测不同培养条件下A-549细胞中HIF-1α、HIF-2α的表达水平。结果 HIF-1α、HIF-2α蛋白的表达在常氧状态下较低,在低氧状态下明显升高,差异具有统计学意义;氧浓度越低,HIF-1α、HIF-2α蛋白的表达水平越高,差异均具有统计学意义;HIF-1α蛋白的表达在低氧2 h即升高,8 h出现表达高峰期,8～16 h出现平台期,32 h出现表达下降,差异具有统计学意义;HIF-2α蛋白随低氧时间的延长,表达逐渐增强。结论在低氧状态下,HIF-1α、HIF-2α的表达均升高,HIF-2α的表达具有时序性变化规律,可能具有更加重要的生物学意义。
Objective To investigate the changes of hypoxia-inducible factor(HIF-1α, HIF-2α) expression level in lung cancer A-549 cells under normoxic conditions, different hypoxia durations, and different oxygen concentrations. Methods A549 cells were divided into normoxic group, time control group, and oxygen concentration control group. Western blot was used to detect the expression of HIF-1α and HIF-2α in A-549 cells.Results The expression of HIF-1α and HIF-2α protein were lower under normoxia and significantly increased under hypoxic conditions. The difference was statistically significant. The lower the oxygen concentration, the more HIF-1α and HIF-2α protein expression levels were. The differences between high and high were statistically significant. The expression of HIF-1α protein increased at 2 h after hypoxia, peaked at 8 h, appeared plateau at 8 to 16 h,and decreased at 32 h, with a statistically significant difference. HIF-2α proteins gradually increased with prolonged hypoxia. Conclusions Under hypoxic conditions, the expression of HIF-1α and HIF-2α are increased,and the expression of HIF-2α has a time-dependent pattern, which may have more important biological significance.
Objective To investigate the changes of hypoxia-inducible factor(HIF-1α, HIF-2α) expression level in lung cancer A-549 cells under normoxic conditions, different hypoxia durations, and different oxygen concentrations. Methods A549 cells were divided into normoxic group, time control group, and oxygen concentration control group. Western blot was used to detect the expression of HIF-1α and HIF-2α in A-549 cells.Results The expression of HIF-1α and HIF-2α protein were lower under normoxia and significantly increased under hypoxic conditions. The difference was statistically significant. The lower the oxygen concentration, the more HIF-1α and HIF-2α protein expression levels were. The differences between high and high were statistically significant. The expression of HIF-1α protein increased at 2 h after hypoxia, peaked at 8 h, appeared plateau at 8 to 16 h,and decreased at 32 h, with a statistically significant difference. HIF-2α proteins gradually increased with prolonged hypoxia. Conclusions Under hypoxic conditions, the expression of HIF-1α and HIF-2α are increased,and the expression of HIF-2α has a time-dependent pattern, which may have more important biological significance.
引文
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[18]ROSATO A,PIVETTA M,PARENTI A,et al.Survivin in esophageal cancer:An accurate prognostic marker for squamous cell carcinoma but not adenocarcinoma[J].Int J Cancer,2006,119(7):1717-1722.
[2]秦承东,任正刚,汤钊猷.缺氧微环境在肿瘤进展中的作用[J].肿瘤,2016,36(1):96-102.
[3]URRUTIA A,ARAGONES J.HIF Oxygen sensing pathways in lung biology[J].Biomedicines,2018,6(2):68.
[4]GAO Z J,WANG Y,YUAN W,et al.HIF-2αnot HIF-1αoverexpression confers poor prognosis in non-small cell lung cancer[J].Tumor Biology,2017,39(6):1010428317709637.
[5]陈万青,孙可欣,郑荣寿,等.2014年中国分地区恶性肿瘤发病和死亡分析[J].中国肿瘤,2018,27(1):1-14.
[6]HERBST R S,MORGENSZTERN D,BOSHOFF C.The biology and management of non-small cell lung cancer[J].Nature,2018,553(7689):446.
[7]VAN ELMPT W,ZEGERS C M L,REYMEN B,et al.Multiparametric imaging of patient and tumour heterogeneity in nonsmall-cell lung cancer:quantification of tumour hypoxia,metabolism and perfusion[J].Eur J Nucl Med Mol Imaging,2016,43(2):240-248.
[8]HSU Y L,HUNG J Y,CHANG W A,et al.Hypoxic lung cancer-secreted exosomal miR-23a increased angiogenesis and vascular permeability by targeting prolyl hydroxylase and tight junction protein ZO-1[J].Oncogene,2017,36(34):4929.
[9]SCHITO L,SEMENZA G L.Hypoxia-inducible factors:master regulators of cancer progression[J].Trends in cancer,2016,2(12):758-770.
[10]RIVERA L B,BERGERS G.Intertwined regulation of angiogenesis and immunity by myeloid cells[J].Trends in immunol,2015,36(4):240-249.
[11]BISHOP T,RATCLIFFE P J.HIF hydroxylase pathways in cardiovascular physiology and medicine[J].Circ Res,2015,117(1):65-79.
[12]江丽霞,施少华,施桥发,等.缺氧诱导因子HIF-1α与HIF-2α在宫颈癌组织中的表达及相关性研究[J].实用医学杂志,2012,28(12):1942-1943.
[13]ROMERO C M,WANG H,BRUICK R.Selective regulation of hypoxia inducible factors by endogenous ligands[J].FASEB J,2017,31(1_supplement):908.10-908.10.
[14]PALAZON A,GOLDRATH A W,NIZET V,et al.HIF transcription factors,inflammation,and immunity[J].Immunity,2014,41(4):518-528.
[15]DEVRAJ G,BEERLAGE C,BRUNE B,et al.Hypoxia and HIF-1 activation in bacterial infections[J].Microbes Infect,2017,19(3):144-156.
[16]KAPITSNOU P P,RAJENDRAN G,ASTLEFORD L,et al.The endothelial PHD2/HIF-2 axis regulates pulmonary artery pressure in mice[J].Mol Cell Biol,2016,36(10):1584-1594.
[17]UCHIDA T,ROSSIGNOL F,MATTHAY M A,et al.Prolonged hypoxia differentially regulates hypoxia-inducible factor(HIF)-1αand HIF-2αexpression in lung epithelial cells IM-PLICATION OF NATURAL ANTISENSE HIF-1α[J].J Biolog Chem,2004,279(15):14871-14878.
[18]ROSATO A,PIVETTA M,PARENTI A,et al.Survivin in esophageal cancer:An accurate prognostic marker for squamous cell carcinoma but not adenocarcinoma[J].Int J Cancer,2006,119(7):1717-1722.