Hsp90抑制剂槚如酸对乳腺癌MDA-MB-231细胞增殖、侵袭、迁移的影响  

Effect of anacardic acid, a Hsp90 inhibitor, on proliferation, invasion and migration of breastcancer MDA-MB-231 cells

作　　者：李红梅[1] 聂丽娟[1] 霍强[1] 赵素容[1] 马涛[1] 吴成柱[1] 刘浩[1] 

机构地区：[1]蚌埠医学院药学系,安徽蚌埠233030

出　　处：《南方医科大学学报》2015年第3期355-359,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81302671;81372899);安徽省自然科学基金(1408085QH162);蚌埠医学院科研基金(Bykf1326)~~

摘　　要：目的探讨热休克蛋白90(Hsp90)抑制剂槚如酸对人乳腺癌MDA-MB-231细胞增殖、侵袭、迁移的影响。方法采用体外malachite green-molybdate显色反应和ATP-琼脂糖凝胶结合实验检测槚如酸对Hsp90抑制活性;MTT法检测药物对MDA-MB-231增殖抑制效应;Transwell小室法检测药物对细胞侵袭和迁移影响;免疫印迹法检测药物对相关蛋白基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制剂-1(TIMP-1)、Hsp90、热休克蛋白70(Hsp70)表达的影响。结果槚如酸为特异性的Hsp90抑制剂,其抑制IC50值为82.5μmol/L。MTT结果显示,槚如酸具有明显抑制细胞增殖的作用,且呈现浓度依赖关系,其IC50值为29.3μmol/L。不同浓度的槚如酸(12.5,25.0,50.0μmol/L)处理36 h后,与对照组相比,药物对细胞侵袭抑制率分别为:23.6%、56.6%、67.0%,P<0.05。槚如酸(12.5,25.0,50.0μmol/L)处理24 h后,与对照组相比,药物对细胞迁移抑制率分别为:30.0%、45.5%、77.5%,P<0.05。免疫印迹法结果显示,槚如酸随着浓度的增加,诱导MMP-9降解越明显,而对Hsp90和Hsp70蛋白表达均没有影响。结论槚如酸能明显抑制乳腺癌细胞增殖、侵袭、迁移能力,其机制可能是通过抑制Hsp90 ATPase活性以及导致下游的顾客蛋白的MMP-9表达下调有关。Objective To explore the effect of the Hsp90 inhibitor anacardic acid on cell proliferation, invasion and migration of breast cancer MDA-MB-231 cells. Methods The inhibitory effect of anacardic acid on Hsp90 was assessed with in vitro ATPase inhibition assay and ATP-sepharose binding assay. MTT assay was used to detect the growth inhibition induced by anacardic acid in MDA-MB-231 cells. Transwell assays were used to evaluate MDA-MB-231 cell invasion and migration. Western blotting was performed to assess the effect of anacardic acid in triggering the degradation of MMP- 9, TIMP- 1, Hsp90, and Hsp70.Results Anacardic acid exhibited a modest activity of ATPase inhibition with an IC50 value of 82.5 μmol/L. Anacardic acid significantly suppressed the proliferation of MDA- MB- 231 cells in a dose- dependent manner（IC50value of 29.3 μmol/L）.Treatment with 12.5, 25, and 50 μmol/L anacardic acid for 36 h caused inhibition of cell invasion by 23.6%, 56.6%, and 67.0% in MDA-MB-231 cells, respectively（P〈0.05）, and anacardic acid treatment for 24 h inhibited the cell migration by 30.0%, 45.5%,and 77.5%, respectively（P〈0.05）. Anacardic acid dose-dependently induced MMP-9 degradation, but did not obviously affect Hsp90 or Hsp70 expressions. Conclusion Anacardic acid can significantly inhibit the proliferation, invasion, and migration of MDA-MB-231 cells, the mechanism of which may involve the inhibition of Hsp90 ATPse activity and down-regulation of MMP-9 expression.

关 键 词：槚如酸 热休克蛋白90 三阴性乳腺癌 抗肿瘤活性 

分 类 号：R737.9[医药卫生—肿瘤]