NIBP在结肠癌细胞中通过调控NF-κB信号通路促进MMP2、MMP9表达

NIBP regulates colon cancer cell express MMP2 and MMP9 via NF-κB signaling pathways

目的:观察慢病毒介导的NIBP(NIK and IKKβbinding protein)基因转染结肠癌细胞株HT29后,细胞迁移能力以及细胞内p65、MMP2、MMP9 m RNA和蛋白表达的变化。方法:分为未经转染的HT29细胞(HT29组)、转染空载的HT29细胞(HT29-NC组)和转染NIBP的HT29细胞(HT29-NIBP稳转组)。采用Transwell试验检测细胞迁移能力;Q-PCR法检测NIBP、p65、MMP2、MMP9的m RNA表达;Western Blot法检测NIBP、p65、磷酸化p65(p-p65)的蛋白表达;ELISA法检测MMP2、MMP9的分泌。结果:高表达NIBP能增强结肠癌细胞株HT29的迁移能力,并主要通过增加p-p65从而促进MMP2、MMP9 m RNA及蛋白表达(P<0.05)。结论:NIBP可能通过激活NF-κB信号通路促进结肠癌细胞分泌MMP-2、MMP-9,从而促进结肠癌细胞的侵袭转移。

Objective To observe after the NIBP （NIK and IKK beta binding protein） gene transfected into colon cancer cell lines HT29, the migration of cells and expression of p65, MMP2, MMP9 mRNA and proteins. Methods The experimental group： divided into without transfection HT29 cell （HT29 group） and transfection no-load HT29 cell （HT29-NC group） and transfection NIBP HT29 cell （HT29-NIBP steady group）. Using the Transwell test to detect cell migration ability. Q-PCR method to detect the mRNA expressions of NIBP, p65, MMP2, MMP9. Western Blot method to detect the expressions of NIBP, p65, phosphorylation p65 （p-p65） proteins. The method of ELISA was used to detect the secretion of matrix metalloproteinase （MMP）-2, MMP-9. Results The high expression of NIBP may enhance the migration ability of colon cancer cell lines HT29, increasing the expression of p-p65, MMP2, MMP9 mRNA and proteins （P 〈 0.05）. Conclusion NIBP potently enhances colon cancer HT29 cell migration and invasion. Activation of NF-κB signaling pathways resulted in up-regulation of MMP-2 and MMP-9 maybe one of its molecular mechanisms.