文章摘要
王耀辉,马〓娟,束〓为,彭〓韬,张〓海,白小明,张〓丽,冷〓静.PGE2通过cAMP-PKA信号转导通路促进肝癌细胞VEGF表达[J].南京医科大学学报,2010,(3):299~303
PGE2通过cAMP-PKA信号转导通路促进肝癌细胞VEGF表达
Prostaglandin E2 increased the expression of vascular endothelial growth factor (VEGF) by activation of cAMP-PKA pathway in hepatocellular carcinoma cells
投稿时间:2009-10-13  
DOI:10.7655
中文关键词: 肝细胞癌  PGE2  EP受体  VEGF  cAMP-PKA
英文关键词: hepatocellular carcinoma  PGE2  EP receptor  VEGF  cAMP-PKA
基金项目:国家自然科学基金资助项目(30470784,30871015)
作者单位
王耀辉 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
马〓娟 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
束〓为 口腔医学研究所,南京医科大学附属口腔医院牙周科,江苏 南京〓210029 
彭〓韬 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
张〓海 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
白小明 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
张〓丽 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
冷〓静 南京医科大学生殖医学重点实验室,肿瘤中心,病理学系,江苏 南京〓210029 
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中文摘要:
      目的:探讨PGE2对人肝癌细胞HuH7 VEGF表达的影响及其可能涉及的信号转导通路?方法:用PGE2?EP1-4四种受体激动剂?SQ22536+PGE2?H89+PGE2处理HuH7细胞,Real-time PCR?Western blot?ELISA等检测VEGFmRNA?蛋白表达的变化?结果:10 μmol/L PGE2处理HuH7细胞24 h后,VEGFmRNA的表达水平上升了264%(P < 0.01)?细胞培养上清中VEGF蛋白表达水平上升了24%(P < 0.01);10 μmol/L PGE2处理HuH7细胞6?12?24 h后,胞浆中VEGF蛋白的表达水平分别上升了13.6%?25.7%?39.6% (P < 0.01);10 μmol/L EP2受体激动剂处理HuH7细胞24 h后,胞浆?细胞培养上清中VEGF蛋白表达水平与对照组相比分别上升了32.3%?20.1%(P < 0.01),而10 μmol/L EP1?EP3?EP4受体激动剂处理HuH7细胞24 h后,VEGF的表达水平无明显改变;30 μmol/L PKA抑制剂(H89)?500 μmol/L腺苷酸环化酶抑制剂(SQ22536)处理HuH7细胞24 h后,胞浆VEGF的表达水平分别为对照组的91.6%?89.6%,细胞培养上清中VEGF的表达水平分别为对照组的98.3%?91.9%,与PGE2组相比均有统计学差异(P < 0.01)?结论:PGE2可上调HuH7细胞VEGF的表达,其作用机制可能是通过EP2受体激活cAMP-PKA信号转导通路所致?
英文摘要:
      Objective:To explore VEGF expression induced by PGE2 in human hepatocellular carcinoma cells HuH7 and the possible mechanisms of signal transduction pathways. Methods:HuH7 cells were treated with PGE2,EP1-4 receptor agonist,SQ22536+PGE2,H89+PGE2. VEGF mRNA and protein expression were examined by Real-time PCR,Western blot and ELISA in HuH7 cells. Results:The expression of VEGF mRNA in HuH7 cells were increased by 264%(P < 0.01),and the expression of VEGF in culture supernatant of HuH7 cells were increased by 24%(P < 0.01)after treated with PGE2(10 μmol/L) for 24 h. The expression of VEGF in cytoplasm of HuH7 cells were increased by 13.6%,25.7%,39.6% (P < 0.01)after treated with PGE2 (10 μmol/L) for 6 h,12 h,24 h respectively. The expression of VEGF in cytoplasm and culture supernatant of HuH7 cells were increased by 32.3% and 20.1%(P < 0.01)after treated with EP2 receptor agonist(10 μmol/L) for 24 h respectively,but have no changes after treated with EP1,EP3,EP4(10 μmol/L) receptor agonist. After the treatment of H89 (30 μmol/L) and SQ22536(500 μmol/L) for 24 h,the expression of VEGF in cytoplasm of HuH7 cells was decreased by 91.6% and 89.6%,and the expression of VEGF in culture supernatant of HuH7 cells was decreased by 98.3% and 91.9%,respectively,vs PGE2(P < 0.01). Conclusion:It is suggested that PGE2 might up-regulate the expression of VEGF through EP2 receptor of HuH7 cells,which could be partly related to the cAMP-PKA signal conduction pathway.
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