Effects Of Inflammatory Chemokines CCL1, CCL20 And CXCL5 On Key Protein Phosphorylation In Hepatocellular Carcinoma Cells

Objective: Primary hepatic carcinoma(PHC)is one of the common malignant tumors.Studies have shown that inflammation plays an important role in the development of hepatocellular carcinoma.In recent years,it has been found that JAK/STAT and ERK/p38 signaling pathways are involved in tumor cell growth,proliferation,differentiation,metastasis and apoptosis.The development of malignant tumors is related to the activation of related signaling pathways.It is found that some inflammatory chemokines participate in the germinal process of PHC by activating related signaling pathways,and are closely related to the development and prognosis of liver cancer.The aim of this study was to investigate the effects of inflammatory chemokines on phosphorylation of STAT3,ERK,p38,JNK,and NF-κB proteins in hepatocellular carcinoma cells,and hope to further understand the mechanism of inflammation leading to the development of hepatocellular carcinoma.Methods:(1)Two human hepatoma cell lines SK-Hep1 and Huh7 were selected,and two cells were cultured respectively.(2)Hepatoma cells were starved for 24 hours in serum-free medium,and then the total protein of those cells were extracted after hepatoma cells were treated with CCL1,CCL20 and CXCL5 for 0 min,5 min,15 min,30 min,60 min,and 120 min,respectively;(3)Western-blot method was used to detect the phosphorylation level of STAT3,p38,ERK,NF-κB,and JNK proteins in the above cellular proteins,and to identify the time point of the highest level of phosphorylated protein expression.Results:(1)After adding CCL1 to the medium,the phosphorylation levels of STAT3,ERK,p38,JNK,and NF-κB proteins were elevated in SK-Hep1 cells;while only the phosphorylation level of STAT3 was changed in Huh7 cells.(2)After adding CXCL5,the phosphorylation levels of STAT3,p38,JNK and NF-κB proteins were up-regulated inHep1 cells;while phosphorylation of STAT3,ERK,p38 and NF-κB was up-regulated in Huh7 cells.(3)Phosphorylation levels of STAT3,ERK,JNK,and NF-κB were altered in Hep1 cells after stimulation with CCL20;whereas,only phosphorylation of STAT3,ERK,and NF-κB was up-regulated in Huh7 cells.Conclusion: The inflammatory chemokines CCL1,CCL20 and CXCL5 can regulate the phosphorylation of STAT3,ERK,p38,JNK and NF-κB proteins in hepatocellular carcinoma cells.The results help to further elucidate the cellular molecular mechanisms by which inflammation leads to the development of hepatocellular carcinoma.