蒙花苷通过调控IKK/NF-κB信号通路抑制人乳腺癌MDA-MB-231细胞的迁移和侵袭

Linarin inhibits migration and invasion abilities of human breast cancer MDA-MB-231 cells through IKK/NF-κB signaling pathway

目的:探究蒙花苷(LIN)对人乳腺癌MDA-MB-231细胞迁移和侵袭能力的影响,并探讨可能的分子机制。方法:用不同浓度(5、10、20、40、80和160μmol/L)的LIN处理MCF-7、MDA-MB-231和MCF-10A细胞24 h后,CCK-8法和集落形成实验检测细胞的增殖能力,Transwell法检测细胞的迁移和侵袭能力,Western blot检测Snail、E-cadherin、基质金属蛋白酶9(MMP-9)、NF-κB抑制蛋白α(IκBα)、磷酸化IκB激酶α/β(p-IKKα/β)和磷酸化p65(p-p65)的蛋白水平。结果:LIN可剂量依赖性地抑制MDA-MB-231细胞活力(P<0.05),IC 50为55.89μmol/L;20μmol/L LIN作用24 h后,MDA-MB-231细胞集落形成率显著降低(P<0.05);与对照组相比,5μmol/L和10μmol/L LIN作用24 h后,MDA-MB-231细胞迁移率和侵袭率显著降低(P<0.05),Snail和MMP-9的蛋白水平下调(P<0.05),E-cadherin的蛋白水平上调(P<0.05),IKKα/β和p65蛋白的磷酸化水平降低,IκBα的蛋白水平上调(P<0.05);同时,IKK-16(IKKα/β抑制剂)和PDTC(NF-κB抑制剂)也可下调MDA-MB-231细胞中Snail和MMP-9的蛋白水平(P<0.05),并上调E-cadherin的蛋白水平(P<0.05)。结论:LIN可能通过抑制IKK/NF-κB信号通路活化而下调Snail和MMP-9蛋白水平并上调E-cadherin蛋白水平,最终导致MDA-MB-231细胞迁移和侵袭能力降低。

AIM:To investigate the effect of linarin(LIN)on the migration and invasion abilities of human breast cancer MDA-MB-231 cells and its underlying mechanism.METHODS:MCF-7,MDA-MB-231 and MCF-10A cells were cultured in vitro and treated with LIN at 5,10,20,40,80 and 160μmol/L for 24 h,and the cell proliferation was measured by CCK-8 assay and colony formation assay.The protein levels of Snail,E-cadherin,matrix metalloproteinase-9(MMP-9),IκBα,p-IKKα/βand p-p65 were determined by Western blot.RESULTS:LIN remarkably reduced the viability of MDA-MB-231 cells in a dose-dependent manner(P<0.05),and the IC 50 was 55.89μmol/L for 24 h.LIN decreased the colony formation rate of MDA-MB-231 cells at the concentration of 20μmol/L(P<0.05).After exposed to LIN at 5μmol/L and 10μmol/L for 24 h,the migration and invasion abilities of the MDA-MB-231 cells were significantly reduced(P<0.05),the protein expression levels of E-cadherin and IκBαwere up-regulated(P<0.05),the protein expression levels of Snail and MMP-9 were down-regulated(P<0.05),and the phosphorylation levels of IKKα/βand p65 were decreased(P<0.05)in comparison with the control group.Meanwhile,IKK-16(IKKα/βinhibitor)and PDTC(NF-κB inhibitor)also down-regulated the protein expression levels of Snail and MMP-9(P<0.05),and up-regulated the protein expression level of E-cadherin(P<0.05).CONCLUSION:LIN down-regulates the protein expression levels of Snail and MMP-9,and up-regulates the protein expression level of E-cadherin most likely through inhibiting IKK/NF-κB signaling pathway,and ultimately lead to decreases in the migration and invasion abilities of MDA-MB-231 cells.