17-AAG通过STAT3通路调控胃癌细胞VEGF基因及蛋白表达作用探讨  

Regulatory effect of 17-AAG on VEGF expression in gastric cancer cells via STAT3 signaling pathway

作　　者：赵恩宏[1] 许倩[2] 肖丽君[3] 刘镭[3] 赵爽[3] 高亚贤[3] 郑鑫[1] 

机构地区：[1]承德医学院附属医院肿瘤外一科,河北省承德市067000 [2]承德医学院基础医学研究所 [3]承德医学院免疫教研室

出　　处：《中国肿瘤临床》2013年第1期16-20,28,共6页Chinese Journal of Clinical Oncology

基　　金：河北省科技厅科学技术项目(编号:072761450);河北省教育厅自然科学项目(编号:2007326);河北省人口和计划生育委员会科技项目(编号:2011-A15)资助

摘　　要：目的:观察17-烯丙胺-17-脱甲氧格尔德霉素(17-allylamino-17-desmethoxy-geldanamycin,17-AAG)对人胃癌MKN-45细胞信号转导和转录活化蛋白3(signal transducer and activator of transcriptions 3,STAT3)、血管内皮生长因子(vascularendothelial growth factor,VEGF)mRNA及蛋白表达水平的影响,并探寻STAT3通路所发挥的作用。方法:体外培养人胃癌MKN-45细胞,分别给予不同剂量及不同作用时间17-AAG进行作用,四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖能力;RT-PCR法和Western Blotting法检测各组细胞STAT3和VEGF mRNA和蛋白的表达水平。结果:0.165～10 mg/L的17-AAG作用24、48 h后对MKN-45细胞有显著的抑制作用,且有明显的时间、剂量依赖性;1.0、2.0、3.0、5.0 mg/L的17-AAG作用48 h后,各组细胞STAT3和VEGF mRNA和蛋白表达均下调,且具有浓度依赖性;3.0 mg/L 17-AAG作用12、24、48h后,各组细胞STAT3和VEGF mRNA和蛋白表达均下调,且具有时间依赖性。结论:17-AAG对人胃癌MKN-45细胞的增殖具有明显的抑制作用,并能抑制STAT3和VEGF mRNA和蛋白的表达,17-AAG可能通过对STAT3通路的负性调控作用而抑制VEGF表达。Objective： This study aims to investigate the effects of 17-allylamino -17-desmethoxy-geldanamycin （17-AAG） on the gene and protein expressions of VEGF and STAT3 pathways in MKN-45 human gastric cancer cells, and explore the potential mechanisms of the STAT3 signaling pathway that mediate these effects. Methods： MKN-45 cells were seeded in Dulbecco＇s Modified Eagle Medium. The thiazolyl blue method, i. e., methyl thiazolyl tetrazolium assay, was performed to evaluate the inhibitory effects of 17-AAG on the proliferation of MKN-45 cells at different times, with different doses. The gene and protein expression of VEGF and STAT3 were determined by reverse transcription polymerase chain reaction （RT - PCR） and Western blot analysis. Results： MKN-45 cells were treated with 17-AAG at 0.165 mg/L to 10 mg/L for 24 and 48 h, respectively. 17-AAG significantly inhibited MKN-45 cell proliferation in a time- and dose-dependent manner. The results of the RT - PCR and Western blot assays showed that the gene and protein expressions of VEGF and STAT3 in MKN-45 cells induced by 17-AAG were significantly down-regulated in a dose-dependent manner when the cells were treated with 17-AAG at 1.0, 2.0, 3.0, and 5.0 mg/L for 48 h. The gene and protein expressions of VEGF and STAT3 in the MKN-45 cells were significantly down-regulated in a time-dependent manner when the cells were treated with 17-AAG at 3.0 mg/L for 12, 24, and 48 h. Conclusion： The drug 17-AAG can inhibit cell proliferation and down-regulate the gene and protein expressions of VEGF and STAT3 in MKN-45 human gastric cancer cells in vitro; the down-regulation of VEGF expression may be associated with the STAT3 signaling pathway.

关 键 词：17-AAG细胞增殖STAT3 VEGF 

分 类 号：R735.2[医药卫生—肿瘤]