CXCL1促进内质网应激在肝癌肿瘤微环境中传递

CXCL1 Promotes the Transmission of Endoplasmic Reticulum Stress Between Cells in the Microenvironment in Hepatocellular Carcinoma

目的:研究趋化因子1(CXCL1)对肝癌肿瘤微环境中肿瘤细胞间及肿瘤细胞和巨噬细胞间内质网应激(ERS)的作用。方法:收集受衣霉素(TM)刺激的HepG2细胞的上清作为条件培养基,用于培养未受TM刺激的HepG2和THP-1细胞,检测培养基中未折叠蛋白反应(UPR)相关蛋白IRE1、PERK和ATF6的mRNA表达,构建ERS在细胞间传递的模型;合成si-CXCL1,瞬时转染HepG2细胞,RT-PCR检测HepG2细胞的CXCL1的转染效率,ELISA检测上清中CXCL1的表达水平;TM刺激CXCL1敲低的HepG2细胞,收集其上清用于培养未受TM刺激的HepG2和TPH-1细胞,RT-PCR检测HepG2细胞和THP-1细胞中UPR相关的IRE1、PERK及ATF6的mRNA表达。结果:用TM刺激后的条件培养基培养HepG2和TPH-1细胞时,2种细胞中的IRE1、PERK、ATF6表达均上调,构建了ERS在细胞间传递的模型;si-CXCL1转染HepG2细胞后,与对照组相比,si-CXCL1组HepG2细胞中CXCL1的表达降低,同时ELISA检测培养基中的CXCL1也降低;用TM刺激CXCL1敲低的HepG2细胞的上清培养未受TM刺激的HepG2和TPH-1细胞后,与对照组相比,CXCL1血清敲低组培养的细胞中IRE1、PERK和ATF6的mRNA水平均下降。结论:CXCL1能够促进肝癌肿瘤微环境中内质网应激在细胞间传递。

Objective:To investigate the role of CXC chemokine ligand 1(CXCL1)in the transmission of endoplasmic reticulum stress(ERS)in hepatocellular carcinoma.Methods:Firstly,a model of transmission of ERS by CXCL1 was established to test the feasibility of signal transmission in HepG2 cells as well as HepG2 and THP-1 cells.The cultured serum of tunicamycin(TM)-stimulated HepG2 cells was collected and added into the medium of unstimulated HepG2 and THP-1 cells.RT-PCR was used to test the relative mRNA level of three unfolded protein reaction(UPR)related protein,IRE1,PERK and ATF6.Secondly,si-CXCL1 was used to analyze the effect of CXCL1 in ERS transmission.Results:A model of transmission of ERS was successfully constructed,notably,CXCL1 siRNA cells showed reduced ability in inducing mRNA expression of IRE1,PERK and ATF6 in both HepG2 and THP-1 cells.Conclusion:CXCL1 promotes the transmission of ERS between cells in the microenvironment in hepatocellular carcinoma.