17-烯丙氨基格尔德霉素对人乳腺癌细胞基质金属蛋白酶表达调控作用的研究  

Effects of 17-AAG on the Expression of MMP in Human Breast Neoplasm MCF-7 Cells

作　　者：许倩[1] 肖丽君[2] 赵恩宏[3] 魏宪嘉[2] 张海燕[2] 赵爽[2] 郑鑫[3] 

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

出　　处：《中国全科医学》2013年第3期271-274,共4页Chinese General Practice

基　　金：河北省科技厅科学技术项目(072761450);河北省教育厅自然科学项目(2007326);河北省人口和计划生育委员会科技项目(2011-A15);承德医学院自然科学项目(201204)

摘　　要：目的观察17-烯丙氨基格尔德霉素(17-AAG)对人乳腺癌MCF-7细胞基质金属蛋白酶(MMP)-2、MMP-9 mRNA及其表达的调控作用。方法体外培养人乳腺癌MCF-7细胞,分别给予不同剂量及不同作用时间的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;反转录聚合酶链反应(RT-PCR)法和蛋白印迹(Western Blotting)法检测MCF-7细胞的MMP-2、MMP-9 mRNA和蛋白表达水平。结果 (1)10.000、5.000、2.500、1.250、0.625、0.310、0.165 mg/L 17-AAG及空白对照组培养24 h和48 h时A值比较,差异均有统计学意义(P<0.05)。体外作用24 h后IC50均值为34.61,体外作用48 h的IC50均值为6.22。(2)RT-PCR法检测不同浓度17-AAG对MCF-7细胞MMP-2、MMP-9 mRNA表达的影响,结果显示,对照组及1.00 mg/L组、2.00 mg/L组、3.00 mg/L组、5.00 mg/L组MMP-2/β-actin和MMP-9/β-actin比较,差异均有统计学意义(P<0.05);其中各组间两两比较,差异均有统计学意义(P<0.05)。(3)Western Blotting法检测不同浓度17-AAG对MCF-7细胞MMP-2、MMP-9表达的影响,结果显示,对照组及1.00 mg/L组、2.00 mg/L组、3.00mg/L组、5.00 mg/L组MMP-2/β-actin和MMP-9/β-actin比较,差异均有统计学意义(P<0.05);其中各组间两两比较,差异均有统计学意义(P<0.05)。结论 17-AAG对人乳腺癌MCF-7细胞的增殖具有明显的抑制作用,并能抑制MMP-2和MMP-9的表达,具有剂量依赖性。Objective To investigate the effects of 17-AAG on the genes and proteins expression of MMP-2 and MMP-9 in human breast neoplasm MCF-7 cells. Methods MCF-7 cells were cultivated in DMEM medium,and MTT method was used to evaluate the inhibitory effects of 17-AAG on the proliferation of MCF-7 cells at different time and different dose.The genes and proteins expression of MMP-2 and MMP-9 were detected by RT-PCR and Western Blotting analysis. Results（1） The A value of the control group and groups treated by 17-AAG of 10.000,5.000,2.500,1.250,0.625,0.310 and 0.165 mg/L for 24 h and 48 h all showed statistically significant differences（P0.05）. The levels of IC50 treated by different concentration of 17-AAG for 24 h are 34.61,the 48 h′s are 6.22.（2） RT-PCR method was used to detect the influence of 17-AAG with different concentration on the expression of MMP-2 and MMP-9 mRNA in MCF-7 cells,and the results showed that MMP-2/β-actin and MMP-9/β-actin all showed statistically significant differences among control group,1.00 mg/L group,2.00 mg/L group,3.00 mg/L group and 5.00 mg/L group（P0.05）,and there were also statistically significant differences pairwise（P0.05）.（3） Western Blotting method was used to detect the influence of 17-AAG with different concentration on the expression of MMP-2 and MMP-9 protein in MCF-7 cells,and the results showed that MMP-2/β-actin and MMP-9/β-actin all showed statistically significant differences among control group,1.00 mg/L group,2.00 mg/L group,3.00 mg/L group and 5.00 mg/L group（P0.05）,and there were also statistically significant differences pairwise（P0.05）. Conclusion 17-AAG can inhibit the cell proliferation and down regulate the genes and proteins expression of MMP-2 and MMP-9 of human breast neoplasm MCF-7 cells in a dose-dependent manner.

关 键 词：乳腺肿瘤 17-烯丙氨基格尔德霉素 基质金属蛋白酶2 基质金属蛋白酶9 

分 类 号：R737.9[医药卫生—肿瘤]