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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

李伯和, 袁磊^*

漯河医学高等专科学校分子医学实验室，漯河 462002

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，10~50 μmol/L辣椒碱对MDA-MB-231细胞存活率无显著影响，100~500 μmol/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50 μmol/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。

关键词： 辣椒碱; 乳腺癌 ; 迁移 ; 侵袭 ; MDA-MB-231细胞

分类号：R737.9

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

LI Bai-He, YUAN Lei^*

Laboratory of Molecular Biology, Luohe Medical College, Luohe 462002, China

Abstract

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-Src, 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.

Key words: capsaicin; breast cancer ; migration ; invasion ; MDA-MB-231 cells

收稿日期：2016-08-22　　录用日期：2016-11-01

通讯作者：袁磊　　E-mail: fzyx_yl@163.com

引用本文：

李伯和, 袁磊. 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[J]. 生理学报 2017; 69 (2): 183-188.

LI Bai-He, YUAN Lei. Inhibitory effects of capsaicin on migration and invasion of breast cancer MDA-MB-231 cells and its mechanism. Acta Physiol Sin 2017; 69 (2): 183-188 (in Chinese with English abstract).