趋化因子CXCL1通过整合素β1调控胃癌转移的机制研究

CXCL1 promoting the metastases of gastric cancer via integrinβ1

目的探讨过表达趋化因子CXCL1促进胃癌细胞迁移及相关分子机制。方法应用胃癌细胞株(SGC-7901、BGC-823),重组慢病毒方法构建过表达CXCL1细胞株,siRNA敲低胃癌细胞表达整合素β1(integrinβ1)。Western blotting法检测CXCL1、integrinβ1、基质金属蛋白酶(MMP)-2、MMP-9、FAK、SRC和ERK的表达,Transwell实验评估胃癌细胞的迁移能力。结果过表达CXCL1的SGC-7901和BGC-823细胞中integrinβ1表达水平高于对照细胞,siRNA干扰CXCL1表达后,胃癌细胞integrinβ1表达水平下降。外源性CXCL1分别增强SGC-7901和BGC-823细胞迁移能力(2.40±0.44)倍(P=0.002)和(2.08±0.30)倍(P=0.001)。此外,CXCL1还增强FAK、SRC和ERK的磷酸化水平。integrinβ1-siRNA可以阻断CXCL1所引起的SGC-7901和BGC-823细胞迁移能力增强(P<0.05)及FAK、SRC和ERK的磷酸化水平。CXCL1调控SGC-7901和BGC-823细胞的MMP-2、MMP-9表达,而敲低integrinβ1后MMP-2、MMP-9表达随之下调。MMP抑制剂GM6001抑制7901-CXCL1和823-CXCL1细胞的迁移能力(P<0.05)。结论CXCL1通过integrinβ1激活FAK-SRC-ERK通路和调控MMP-2、MMP-9的表达,最终调控胃癌细胞迁移。

Objective To investigate the effect of CXCL1 on the migration of gastric cancer(GC)cells and its potential mechanism.Methods Lentivirus particles expressing CXCL1 were constructed to establish a stable CXCL1 overexpressing SGC-7901 cell line(7901-CXCL1-GFP)and BGC-823 cell line(823-CXCL1-GFP).siRNA was utilized to block the expression of CXCL1 and integrinβ1 in SGC-7901 and BGC-823 cells.Western blotting was performed to analyze the expression of CXCL1,integrinβ1,FAK,SRC,ERK,MMP-2 and MMP-9.Transwell assays were adopted to evaluate the migration of gastric cancer cells.Results The expression of integrinβ1 was higher in 7901-CXCL1-GFP and 823-CXCL1-GFP cells than that in 7901-GFP and 823-GFP cells.Integrinβ1 expression was down-regulated by blocking CXCL1 with siRNA.Blocking integrinβ1 by siRNA could inhibit CXCL1-induced migration of SGC-7901 and BGC-823 cells.CXCL1 increased the expression of p-FAK,p-SRC and p-ERK in GC cells,which could be blocked by integrinβ1 knockdown with siRNA.CXCL1 induced the expression of MMP-2 and MMP-9 in GC cells through integrinβ1 signaling,meanwhile knockdown of integrinβ1 inhibited the MMP-2 and MMP-9 expression.GM6001,one MMPs inhibitor,significantly reduced CXCL1-induced migration of 7901-CXCL1-GFP and 823-CXCL1-GFP cells(P<0.05).Conclusion Overexpressed CXCL1 promotes migration of GC cells by activating FAK-SRC-ERK pathway and regulating the expression of MMP-2,MMP-9 through integrinβ1 signaling.