毛钩藤碱对缺氧乳腺癌MCF-7细胞迁移和侵袭能力的影响  

Effect of hirsutine on hypoxia-induced migration and invasion abilities in human breast cancer MCF-7 cells

作　　者：翟娜娜[1] 黄器伟[1] 陈奎生[2] 

机构地区：[1]黄河科技学院,河南郑州450099 [2]郑州大学第一附属医院,河南郑州450052

出　　处：《中国病理生理杂志》2017年第11期2009-2014,共6页Chinese Journal of Pathophysiology

基　　金：河南省科技厅科技发展计划(No.122300410434)

摘　　要：目的:观察毛钩藤碱对缺氧诱导的人乳腺癌MCF-7细胞迁移和侵袭能力的影响,并探讨其分子机制。方法:选取MCF-7细胞作为研究对象,采用CCK-8法检测毛钩藤碱的细胞毒性作用;采用细胞划痕实验测量细胞迁移率;采用Transwell侵袭实验检测细胞侵袭力;采用Western blot法检测缺氧诱导因子1α(hypoxia-inducible factor-1α,HIF-1α)、Snail、E-钙黏蛋白(E-cadherin)和基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)的蛋白表达水平;采用RT-PCR法检测HIF-1α的mRNA水平。结果:自32μmol/L开始,随着毛钩藤碱浓度的升高,MCF-7细胞存活率明显降低(P<0.05),IC_(50)为62.82μmol/L;在缺氧条件下,MCF-7细胞的迁移和侵袭能力显著增强(P<0.05),HIF-α、Snail和MMP-9蛋白表达水平明显升高(P<0.05),E-cadherin蛋白表达水平明显下降(P<0.05),HIF-1α的mRNA水平也显著升高(P<0.05);除缺氧MCF-7细胞中HIF-α的mRNA水平未受影响外,上述效应均被毛钩藤碱(16μmol/L)明显抑制(P<0.05)。结论:毛钩藤碱可在体外抑制缺氧诱导的人乳腺癌MCF-7细胞迁移和侵袭,这可能与毛钩藤碱下调HIF-1α、Snail和MMP-9蛋白表达水平,上调E-cadherin蛋白表达水平有关。AIM：To investigate the effect of hirsutine on hypoxia-induced migration and invasion abilities of human breast cancer MCF-7 cells and its possible mechanism.METHODS：CCK-8 assay was employed to detect the cytotoxic effect of hirsutine on the MCF-7 cells.Cell migration was observed by wound healing assay,and cell invasion ability was measured by Transwell invasion assay.Western blot was used to analyze the protein levels of hypoxia-inducible factor-1α（HIF-1α）,Snail,E-cadherin and matrix metalloproteinase-9（MMP-9）.The mRNA levels of HIF-1α was detected by RT-PCR.RESULTS：Hirsutine remarkably reduced the cell viability from 32 μmol/L（P 〈 0.05）,and the IC50 value was62.82 μmol/L.In hypoxia state,MCF-7 cells showed more powerful capabilities of migration and invasion（P 〈 0.05）,higher protein levels of HIF-1α,Snail and MMP-9（P 〈 0.05）,lower protein level of E-cadherin（P 〈 0.05）,and higher mRNA level of HIF-1α（P 〈 0.05）.These hypoxia-induced effects were all inhibited by hirsutine at 16 μmol/L（P 〈0.05）,apart from the mRNA level of HIF-1α.CONCLUSION：Hirsutine inhibits hypoxia-induced migration and invasion in human breast cancer MCF-7 cells most likely via down-regulation of the protein levels of HIF-1α,Snail and MMP-9,and up-regulation of the protein level of E-cadherin.

关 键 词：毛钩藤碱 乳腺癌 缺氧 细胞迁移 细胞侵袭 

分 类 号：R737.9[医药卫生—肿瘤] R285.5[医药卫生—临床医学]