丹参酮ⅡA调节COX-2对人结肠癌HCT-116细胞VEGF表达的影响  

Tan IIA inhibits COX-2-regulated VEGF expression inhuman colon cancer HCT-116 cells

作　　者：周利红[1,2] 王炎[1,2] 范忠泽[1,2] 吴琼[1,2] 刘宣[1,2] 慈书俊[1,2] 李琦[1,2] 

机构地区：[1]上海中医药大学附属普陀医院 [2]上海中医药大学中西医结合肿瘤介入研究所,上海市200062

出　　处：《世界华人消化杂志》2011年第15期1561-1567,共7页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.810729552;上海市科委实验动物专项基金资助项目;No.10140902600;上海市自然基金资助项目;No.09ZR1428500;上海市重点学科基金资助项目;No.S30302;上海市教委科研预算内基金资助项目;No.09JW42~~

摘　　要：目的:研究中药丹参有效活性成分丹参酮ⅡA(TanⅡA)对人结肠癌HCT-116细胞的生长抑制作用及其调控COX-2对HCT-116细胞VEGF表达的影响.方法:利用MTT法检测不同浓度的TanⅡA对人结肠癌HCT-116细胞的生长抑制作用,将pGL3-Basic-COX-2-promoter重组质粒和pRL-TK内参质粒共转染HCT-116细胞,分为空白组、对照组和TanⅡA组.药物作用48h后,双荧光素酶测定法检测COX-2启动子转录活性.将含COX-2CDS序列的pIRESI-COX-2重组质粒转染HCT-116细胞,分为空白组、对照组和TanⅡA组,ELISA法检测细胞培养液中VEGF的表达.结果:TanⅡA对人结肠癌HCT-116细胞有明显的增殖抑制作用,24、48、72h的IC50值分别为40.3μmol/L±5.22μmol/L,12.9μmol/L±3.24μmol/L,8.5μmol/L±1.47μmol/L,24h的最大无毒剂量为10μmol/L.转染pGL3-Basic-COX-2-promoter重组质粒48h后,COX-2启动子活性明显上调(对照组vs空白组,P<0.01),而不同浓度的TanⅡA可明显抑制COX-2启动子活性.转染pIRESI-COX-2重组质粒后,细胞培养液VEGF浓度升高(对照组vs空白组,P<0.05),而10μmol/LTanⅡA处理后,VEGF的表达显著下调(TanⅡA vs对照组,P<0.05).结论:TanⅡA能明显抑制人结直肠癌细胞增殖,并通过调控人结肠癌细胞COX-2表达抑制结直肠癌细胞VEGF的表达.AIM：To investigate the effect of tanshinone IIA（Tan IIA）,an active component of salvia miltior-rhiza,on cell proliferation and cyclooxygenase-2（COX-2）-regulated vascular endothelial growth factor（VEGF）expression in human colon cancer HCT-116 cells.METHODS：MTT assay was used to evaluate the effect of Tan IIA on the proliferation of HCT-116 cells.HCT-116 cells were co-transfected transiently with the recombinant plasmid pGL3-Basic-COX-2-promoter and control vector pRL-TK and randomly divided into blank group, control group,and Tan IIA group.Dual luciferase reporter assay was used to observe the activity of COX-2 promoter 48 h after treatment with Tan IIA.HCT-116 cells were transfected with the pIRESI-COX-2 plasmid and divided into three groups as above.The expression of COX-2 mRNA in transfected cells was detected by realtime PCR,and the level of VEGF in culture medium was assayed by ELISA.RESULTS：Tan IIA had obvious inhibition on the proliferation of HCT-116 cells.The half maximal inhibitory concentrations（IC50）at 24, 48,and 72 h were 40.3μmol/L±5.22μmol/L, 12.9μmol/L±3.24μmol/L,and 8.5μmol/L ±1.47μmol/L,respectively,and the maximal no-effect dose at 24 h was 10μmol/L.At 48 h after transfection with plasmids carrying the COX-2-promoter and COX-2 cDNA,the activity of COX-2 promoter and expression of COX-2 mRNA were obviously up-regulated（both P 〈0.01）,and the concentration of VEGF was three times higher than that in control cells（P〈0.05）. Treatment with 10μmol/L Tan IIA significantly down-regulated the expression of COX-2 and VEGF in transfected cells（P〈0.05）.CONCLUSION：Tan IIA down-regulates VEGF expression and inhibits cell proliferation possibly by regulating COX-2 promoter activity and mRNA expression in HCT-116 cells.

关 键 词：丹参酮ⅡA 环氧合酶2 血管内皮生长因子 结肠癌 

分 类 号：R735.35[医药卫生—肿瘤]