辣椒碱对乳腺癌MDA-MB-231细胞迁移和侵袭的抑制作用及其机制  

Inhibitory effects of capsaicin on migration and invasion of breast cancer MDA-MB-231 cells and its mechanism

作　　者：李伯和[1] 袁磊[1] LI Bai-He YUAN Lei(Laboratory of Molecular Biology, Luohe Medical College, Luohe 462002, Chin)

机构地区：[1]漯河医学高等专科学校分子医学实验室,漯河462002

出　　处：《生理学报》2017年第2期183-188,共6页Acta Physiologica Sinica

基　　金：supported by the Science and Technology Development Project of the Department of Science and Technology of Henan Province;China(No.142102310466);the Natural Science Research Project of Luohe Medical College;Henan Province;China(No.2014-S-LMC09)

摘　　要：本研究旨在观察辣椒碱对人乳腺癌MDA-MB-231细胞迁移和侵袭的影响,并探讨其分子机制。乳腺癌MDA-MB-231细胞经不同浓度辣椒碱作用24 h后进行实验,采用CCK-8法检测细胞活性,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率,采用Western blot检测乳腺癌MDA-MB-231细胞中c-Src、p-c-Src(Tyr416)、黏着斑激酶(focal adhesion kinase,FAK)、p-FAK(Tyr576/577)、桩蛋白(Paxillin)、p-Paxillin(Tyr118)、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)和MMP9蛋白表达水平,采用RT-PCR法检测乳腺癌MDA-MB-231细胞中MMP2和MMP9的mRNA水平。结果显示,10~50μmol/L辣椒碱对MDA-MB-231细胞存活率无显著影响,100~500μmol/L辣椒碱显著降低MDA-MB-231细胞存活率(P<0.05);与正常对照组比较,25和50μmol/L辣椒碱组MDA-MB-231细胞体外迁移率和侵袭率均显著降低(P<0.05),c-Src、FAK和Paxillin蛋白的磷酸化水平均显著下降(P<0.05),MMP2和MMP9的mRNA和蛋白表达水平均下降(P<0.05);并且辣椒碱对MDA-MB-231细胞的上述作用具有剂量依赖性。以上结果提示,低浓度辣椒碱可剂量依赖性地抑制人乳腺癌MDA-MB-231细胞迁移和侵袭,其机制可能涉及辣椒碱对c-Src/FAK/Paxillin信号通路以及MMP2和MMP9表达的抑制。The aim of this study was to investigate the effects of capsaicin on migration and invasion of breast cancer MDA-MB-231 cells and its possible mechanism. The MDA-MB-231 cells were incubated in the medium containing different concentrations of capsaicin for 24 h. CCK-8 assay was employed to detect the cell viability. The cell migration and invasion were assessed by wound healing assay and transwell invasion assay, respectively. The protein levels of c-Sre, p-c-Src （Tyr416）, FAK, p-FAK （Tyr576）, Paxillin, p-Paxillin （Tyr118）, matrix metalloproteinase 2 （MMP2） and MMP9 in the MDA-MB-231 cells were detected by Western blotting. The mRNA expressions of MMP2 and MMP9 were measured by RT-PCR. The result showed that capsaicin （25 and 50 μmol/L） remarkably reduced the abilities of migration and invasion （P 〈 0.05）, inhibited the phosphorylation of c-Src, FAK and Paxillin （P 〈 0.05）, and down-regulated MMP2 and MMP9 expressions at mRNA and protein levels （P 〈 0.05） in MDA-MB-231 cells. These effects of capsaicin were all in dose-dependent manners. These results suggest that capsaicin may suppress the migration and invasion of breast cancer MDA-MB-231 cells by inhibiting the phosphorylations of c-Src, FAK and Paxillin, and down-regulating the mRNA and protein levels of MMP2 and MMP9.

关 键 词：辣椒碱 乳腺癌 迁移 侵袭 MDA-MB-231细胞 

分 类 号：R737.9[医药卫生—肿瘤]