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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 effect of capsaicin on the migration and invasion of human breast cancer MDA-MB-231 cells and to explore its molecular mechanism. Cell proliferation was detected by cell scratch assay. The cell invasion rate was detected by Transwell invasion assay. Western blotting was used to detect the cell invasion rate in breast cancer MDA-MB-231 cells. After treated with different concentrations of capsaicin for 24 hours, the cell viability was measured by CCK- The expression of c-Src, pc-Src (Tyr416), focal adhesion kinase (FAK), p-FAK (Tyr576 / 577), Paxillin, Paxillin (Tyr118), matrix metalloproteinase 2 (MMP2) and MMP9 were detected by RT-PCR. The mRNA levels of MMP2 and MMP9 in breast cancer MDA-MB-231 cells were detected by RT-PCR. The results showed that 10 ~ 50μmol / L capsaicin had no significant effect on the survival rate of MDA-MB-231 cells, 100 ~ 500μmol / L capsaicin significantly reduced the survival rate of MDA-MB-231 cells Compared with the 25 and 50μmol / L capsaicin groups, the migration and invasion rates of MDA-MB-231 cells were significantly decreased (P <0.05), while the phosphorylation levels of c-Src, FAK and Paxillin proteins were significantly decreased (P <0.05 ), MMP2 and MMP9 mRNA and protein expression levels were decreased (P <0.05); and capsaicin on MDA-MB-231 cells in a dose-dependent manner. The above results suggest that low concentration of 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 c-Src / FAK / Paxillin signal pathway and MMP2 and MMP9 expression by capsaicin .