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山东大学学报(医学版)

• 基础医学 • 上一篇    下一篇

缺氧对人胃癌细胞HIF-1α及DEC1表达的影响

胡瑞1,郏雁飞2,郑燕2,马晓丽2,孔毅1,黎娉1 ,汪运山2   

  1. 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
  • 收稿日期:2013-08-27 出版日期:2014-04-10 发布日期:2014-04-10
  • 通讯作者: 汪运山 E-mail:sdjnwys@163.com
  • 基金资助:
    国家自然科学基金(81000869,81272588);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)

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

HU Rui1, JIA Yanfei2, ZHENG Yan2, MA Xiaoli2, KONG Yi1, LI Pin1, WANG Yunshan2   

  1. 1. School of Medicine, Shandong University, Jinan 250012, Shandong, China;
    2. Central Laboratory, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, Shandong, China
  • Received:2013-08-27 Online:2014-04-10 Published:2014-04-10

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24h组、缺氧48h组)、对照组(常氧24h组、常氧48h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。

关键词: ;分化型胚胎软骨发育基因1, 胃肿瘤;缺氧;缺氧诱导因子-1&alpha

Abstract: 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-1α and DEC1 were determined by RT-PCR, Western blotting and immunocytochemistry method, and their correlations were analyzed. Results  The expressions of both mRNA and protein of DEC1 and HIF-1α in BGC823 cell lines were upregulated significantly compared with the control groups under the hypoxia condition (P<0.05). With the extension of hypoxia, the mRNA expressions of HIF-1α and DEC1 increased, while the protein expressions of HIF-1α and DEC1 declined. Immune cytochemistry method also showed that the positive expression rates of HIF-1α and DEC1 in the experimental groups were higher than those in the control groups (P<0.05). Conclusion  Hypoxia can induce the expressions of HIF-1α and DEC1, which have significant correlation, in BGC823 cell lines. During hypoxia, HIF-1α protein 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.

Key words: Gastric neoplasm, Hypoxia inducible factor-1α, Hypoxia, Differentiated embry-ochondrocyte expressed gene 1

中图分类号: 

  • R735.2
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