脂多糖通过诱导NLRP3表达促进膀胱癌细胞侵袭迁移的机制研究

The mechanism of lipopolysaccharide promoting the invasion and migration of bladder cancer cells by inducing the expression of NLRP3

目的采用脂多糖(LPS)处理膀胱癌(BC)5637细胞以模拟炎症微环境,研究炎症对BC细胞侵袭迁移的影响及由NOD样受体热蛋白结构域相关蛋白3(NLRP3)介导的调控机制。方法将5637细胞分为LPS组、MCC950组、LPS/MCC950组和空白对照组4组,采用CCK-8实验检测细胞增殖活力,Transwell侵袭实验检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力,qRT-PCR检测基质金属蛋白酶-2(MMP-2)、钙黏蛋白E(E-cadhjerin)、波形蛋白(Vimentin)、NLRP3 m RNA表达水平,Western blot检测MMP-2、基质金属蛋白酶-9(MMP-9)、E-cadherin、Vimentin、NLRP3蛋白表达水平。结果LPS可显著增强5637细胞的增殖活力,促进其侵袭迁移及上皮间充质转化,并上调NLRP3 mRNA和蛋白的表达;MCC950可显著下调NLRP3蛋白的表达,并抑制LPS促进5637细胞侵袭迁移的效应。结论LPS可通过诱导NLRP3表达来促进BC细胞侵袭迁移,NLRP3炎症小体可能是炎症状态下促进BC进展的重要因子。

Objective In order to study the effects of inflammation on invasion and migration of bladder cancer(BC)cells and the regulatory mechanism mediated by NLRP3,the 5637 BC cells were treated with lipopolysaccharide(LPS)to simulate the inflammatory microenvironment.Methods The 5637 cells were divided into 4 groups as follows:LPS group,MCC950 group,LPS/MCC950 group and blank control group.The proliferation,invasion and migration of cells were detected by CCK-8 assay,Transwell invasion assay and wound healing assay,respectively.Besides,the mRNA expression levels of MMP-2,E-cadherin,Vimentin and NLRP3 were detected by qRT-PCR,and the protein expression levels of MMP-2,MMP-9,E-cadherin,Vimentin and NLRP3 were detected by Western blot.Results LPS could significantly enhance the proliferation activity of 5637 cells,promote their invasion,migration and epithelial-mesenchymal transition,and up-regulate the expressions of NLRP3 mRNA and protein.Conversely,MCC950 could significantly down-regulate the expression of NLRP3 protein and inhibit the positive effect of LPS on invasion and migration of 5637 cells.Conclusion LPS can promote the invasion and migration of BC cells by inducing the expression of NLRP3,and NLRP3 inflammasome may be a critical tumor promotor in BC under the circumstance of inflammation.