缺氧对人胃癌细胞HIF-1α及DEC1表达的影响  

Effect of hypoxia on the expressions of HIF-1α and DEC1 in human gastric cancer cell

作　　者：胡瑞[1] 郏雁飞[2] 郑燕[2] 马晓丽[2] 孔毅[1] 黎娉[1] 汪运山[2] 

机构地区：[1]山东大学医学院,山东济南250012 [2]山东大学附属济南市中心医院中心实验室,山东济南250013

出　　处：《山东大学学报（医学版）》2014年第4期35-38,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(810008869;81272588);国家重点基础研究发展计划(973计划;2012CB966503;2012CB966504);山东省自然科学基金(ZR2012HM061);山东省优秀中青年科学家科研奖励基金(BS2011YY039)

摘　　要：目的研究缺氧对人胃癌细胞株BGC823缺氧诱导因子-1α(HIF-1α)及分化型胚胎软骨发育基因1(DEC1)表达的影响。方法将细胞株BGC823分为实验组(缺氧24 h组、缺氧48 h组)、对照组(常氧24 h组、常氧48 h组)和阴性对照组(PBS作为一抗)。实验组置于5%CO2、0.5%O2缺氧培养箱培养,对照组于37℃、5%CO2的培养箱中培养。用RT-PCR及Western blotting的方法检测两组细胞HIF-1α与DEC1各水平的表达。另外,用免疫细胞化学的方法进一步检测HIF-1α与DEC1的表达并进行相关性分析。结果缺氧处理后,HIF-1α和DEC1的mRNA和蛋白表达水平均明显高于对照组(P<0.05)。HIF-1α及DEC1 mRNA表达水平随缺氧时间增加而上升,HIF-1α及DEC1蛋白表达水平随缺氧时间增加而下降。实验组HIF-1α及DEC1阳性率明显高于对照组(P<0.05)。结论缺氧条件下,胃癌细胞株BGC823中HIF-1α、DEC1各水平的表达增加,二者具有显著相关性。缺氧可能通过诱导HIF-1α的大量表达,转录激活DEC1的表达。DEC1作为HIF-1α的下游靶基因,在胃癌细胞适应缺氧环境中发挥重要作用。Objective To investigate the effect of hypoxia on the expressions of hypoxia inducible factor-1α（ HIF-1α） and differentiated embry-ochondrocyte expressed gene 1 （ DEC1 ） in human gastric cancer cells.Methods The BGC823 cell lines were divided into the experimental groups （ hypoxia for 24 hours group and hypoxia for 48 hours group） , the control groups （ normoxic for 24 hours group and normoxic for 48 hours group） , and the negative control group （ PBS instead of the first antibody） .The experimental groups were cultured in the hypoxia incubator with 5%CO2 and 0.5%O2 , and the control groups were exposed to the normal oxygen condition.The levels of mRNA and protein of HIF-1αand DEC1 were determined by RT-PCR, Western blotting and immunocytochemistry method, and their correla-tions were analyzed.Results The expressions of both mRNA and protein of DEC1 and HIF-1αin BGC823 cell lines were upregulated significantly compared with the control groups under the hypoxia condition （P＆lt;0.05）.With the ex-tension of hypoxia, the mRNA expressions of HIF-1αand DEC1 increased, while the protein expressions of HIF-1αand DEC1 declined.Immune cytochemistry method also showed that the positive expression rates of HIF-1αand DEC1 in the experimental groups were higher than those in the control groups （P＆lt;0.05）.Conclusion Hypoxia can induce the expressions of HIF-1αand DEC1, which have significant correlation, in BGC823 cell lines.During hypoxia, HIF-1αprotein accumulates and activates the transcription of downstream target gene DEC1, which suggests that DEC1 is a marker of activated hypoxic pathways and plays an important role in hypoxia tolerance in gastric cancer.

关 键 词：胃肿瘤 缺氧 缺氧诱导因子-1α 分化型胚胎软骨发育基因1 Hypoxia INDUCIBLE factor-1α DIFFERENTIATED embry-ochondrocyte EXPRESSED gene 1 

分 类 号：R735.2[医药卫生—肿瘤]