HMGB1调控TLR4/MyD88/NF-κB通路对胃癌侵袭及转移的影响

Effect of HMGB1 expression on invasion and migration of gastric cancer cells and TLR4/MyD88/NF-κB signaling pathway

目的探讨高迁移率族蛋白B1(HMGB1)是否可通过激活TLR4/MyD88/NF-κB信号通路促进胃癌(GC)侵袭及转移。方法选取人胃癌细胞株MGC-803、SGC-7901及人胃上皮细胞株GES-1,采用划痕、Transwell实验检测细胞的侵袭及转移,采用Real-time PCR及Western blot检测细胞中HMGB1、TLR4、Myd88、NF-κB p65、p-NF-κB p65的mRNA及蛋白表达水平;选取MGC-803细胞,采用HMGB1 siRNA进行转染,检测细胞恶性行为及TLR4/MyD88/NF-κB通路分子的表达变化。结果与GES-1细胞比较,MGC-803及SGC-7901细胞的迁移及侵袭能力更强(P<0.01),HMGB1、TLR4、Myd88、NF-κB p65 mRNA及蛋白表达水平均明显上调(P<0.01),p-NF-κB p65蛋白表达水平也明显上调(P<0.01);与SGC-7901细胞比较,MGC-803细胞侵袭能力明显增强(P<0.01),HMGB1、TLR4、Myd88、NF-κB p65 mRNA及蛋白的表达水平均明显上调(P<0.01),p-NF-κB p65蛋白表达水平也明显上调(P<0.01);与HMGB1 siRNA NC组比较,HMGB1 siRNA组细胞侵袭及迁移能力均明显下降(P<0.01),TLR 4、Myd 88 mRNA的表达水平均明显下调(P<0.01),p-NF-κB p65蛋白表达水平也明显下调(P<0.01)。结论抑制TLR4/MyD88/NF-κB信号通路的激活及干扰HMGB1的表达,可减轻胃癌细胞株侵袭及转移。

Objective To explore whether high mobility group protein B1(HMGB1)can promote the invasion and metastasis of gastric cancer(GC)by activating TLR4/MyD88/NF-κB signaling pathway.Methods Human gastric cancer cell lines MGC-803,SGC-7901,and human gastric epithelial cell line GES-1 were cultured,and the invasion and metastasis of cells were detected by scratch and Transwell test,The mRNA and protein expression of HMGB1,TLR4,Myd88,NF-κB p65,and p-NF-κB p65 were detected by Real-time PCR and Western blot.The MGC-803 cells were selected and transfected with HMGB1 siRNA.The malignant behavior of the cells and the expression changes of TLR4/MyD88/NF-κB pathway molecules were detected.Results Compared with GES-1,MGC-803,and SGC-7901 had stronger migration and invasion ability(P<0.01),and the expressions of HMGB1,TLR4,Myd88,and NF-κB p65 mRNA and protein were significantly up-regulated(P<0.01),the protein expression of p-NF-κB p65 was significantly up-regulated(P<0.01).Compared with SGC-7901,MGC-803 had better invasion ability(P<0.01),and the expressions of HMGB1,TLR4,Myd88,and NF-κB p65 mRNA and protein were significantly increased(P<0.01),the protein expression of p-NF-κB p65 was significantly up-regulated(P<0.01).Compared with HMGB1 siRNA NC group,the invasion and migration ability of HMGB1 siRNA group cells decreased significantly(P<0.01),and the expressions of TLR4,Myd 88 were down-regulated(P<0.01),and the protein expression of p-NF-κB p65 was significantly down-regulated(P<0.01).Conclusions The inhibition of TLR4/MyD88/NF-κB pathway and HMGB1 expression could suppress the invasion and metastasis of gastric cancer cells.