三氧化二砷对乳腺癌细胞血管内皮生长因子表达的影响  

Effect of arsenic trioxide on the VEGF expression in human breast carcinoma cells line

作　　者：宗建春[1] 吴诚义[1] 吴凯南[1] 

机构地区：[1]重庆医科大学附属第一医院普外科,400016

出　　处：《中华实验外科杂志》2005年第5期532-534,共3页Chinese Journal of Experimental Surgery

基　　金：重庆市卫生局科研项目(03-02-074)

摘　　要：目的研究低氧条件下人乳腺癌细胞株MDA-MB-435S血管内皮生长因子(VEGF)的表达及三氧化二砷(As2O3)对其表达的影响。方法建立MD-MBA-435S细胞体外低氧培养模型。采用逆转录-聚合酶链反应(RT-PCR)、免疫细胞化学染色和Westerblot分别检测0.5、2.0、5.0μmol/LAs2O3对缺氧12h后乳腺癌细胞VEGF基因及蛋白的表达变化;噻唑蓝(MTT)比色法检测不同浓度As2O3对乳腺癌细胞生存的影响。结果高浓度As2O3可以抑制乳腺癌细胞的生存,而低浓度则对此影响较小。缺氧12h后VEGFmRNA和蛋白皆明显增加;3种浓度As2O3对常氧下乳腺癌细胞VEGFmRNA和蛋白无明显影响,而对缺氧12h后细胞VEGFmRNA和蛋白的表达明显抑制,且呈剂量递增效应。结论缺氧可以显著增加乳腺癌细胞VEGF的表达;常氧下As2O3对乳腺癌细胞VEGF的表达无明显抑制,而在低氧下可以明显抑制,其机制可能是通过调控VEGF的上游基因实现的。Objective To investigate the effect of hypoxia on the expression of vascular endothelial growth factor (VEGF) in breast carcinoma cells line (MD-MBA-435S) and the effect of arsenic trioxide (As_2O_3) on the expression of VEGF.Methods Hypoxia culture model in vitro was established.After cultured in hypoxia circumstance for 12 h and intervened by As_2O_3 (0.5 μmol/L, 2.0 μmol/L, 5.0 μmol/L respectively),the VEGF expression in MD-MBA-435S cells line was detected in mRNA level and protein level by RT-PCR,immunocytochemistry and Western blot.The effect of As_2O_3 on viability of the cells was tested by MTT analysis.Results As_2O_3 in 0.5 μmol/L, 2.0 μmol/L had little effect on cell viability,but 5.0 μmol/L As_2O_3 decreased significantly the viability of the cells.After cultured in hypoxia circumstance for 12 h,the VEGF expression was increased significantly in mRNA and protein levels.As_2O_3 at different concentrations had little effect on the VEGF expression in cells cultured under normoxia,but had significant effect on the cells cultured under hypoxia for 12 h.Conclusion VEGF gene and protein can be induced remarkably by hypoxia in breast carcinoma cells line.As_2O_3 had no effect on the VEGF expression in breast carcinoma cell under normoxia,but inhibited significantly the expression of VEGF under hypoxia,which might be contributed to the mechanism by regulating the upstream gene of VEGF.

关 键 词：三氧化二砷 血管内皮生长因子(VEGF) MDA-MB-435S 逆转录-聚合酶链反应 As2O3 免疫细胞化学染色 人乳腺癌细胞株 Wester VEGF基因 mRNA mol/L 低氧条件 培养模型 方法建立 PCR) blot 表达变化 细胞生存 不同浓度 剂量递增 

分 类 号：R737.9[医药卫生—肿瘤]