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

发布时间：2018-02-24 00:34

本文关键词： 辣椒碱乳腺癌迁移侵袭 MDA-MB-细胞　出处：《生理学报》2017年02期 　论文类型：期刊论文

【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(P0.05);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(P0.05),c-Src、FAK和Paxillin蛋白的磷酸化水平均显著下降(P0.05),MMP2和MMP9的mRNA和蛋白表达水平均下降(P0.05);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。
[Abstract]:The purpose of this study was to observe the effect of capsaicin on the migration and invasion of human breast cancer MDA-MB-231 cells, and to explore its molecular mechanism. After 24 h of different concentrations of capsaicin, the cell activity was detected by CCK-8 assay. Cell migration was measured by cell scratch assay, cell invasion rate was detected by Transwell invasion assay, expression levels of MMP9 protein and c-Srcnp-c-Srctr Tyr416 in breast cancer MDA-MB-231 cells, focal adhesion kinase- pFAK tyr57657 / 7, Paxillinp-Paxillin Tyr118, matrix metalloproteinase 2 metalloproteinase 2MMP2) and MMP9 protein expression were detected by Western blot. The mRNA levels of MMP2 and MMP9 in breast cancer MDA-MB-231 cells were detected by RT-PCR assay. The results showed that capsaicin 1050 渭 mol/L had no significant effect on the survival rate of MDA-MB-231 cells. Capsaicin 100 渭 mol/L significantly decreased the survival rate of MDA-MB-231 cells P0.05, and capsaicin 25 and 50 渭 mol/L capsaicin compared with the normal control group. The in vitro migration and invasion of MDA-MB-231 cells in vitro decreased the phosphorylation level of P0.05, c-Srcfak and Paxillin proteins, and the mRNA and protein expression levels of P0.05MMP2 and MMP9 decreased in a dose-dependent manner, and capsaicin had a dose-dependent effect on MDA-MB-231 cells. The above results suggest that, Low concentration capsaicin can inhibit the migration and invasion of human breast cancer MDA-MB-231 cells in a dose-dependent manner. The mechanism may be related to the inhibition of capsaicin on the c-Src / FAK / paxillin signaling pathway and the expression of MMP2 and MMP9.
【作者单位】：漯河医学高等专科学校分子医学实验室;
【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)
【分类号】：R737.9

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