C–C motif chemokine ligand 16 inhibits the progression of liver cirrhosis via inactivating hepatic stellate cells

Background:Liver cirrhosis results from many forms of chronic damage,characterized by accumulation of extracellular matrix.The present study aimed to explore a potential non-invasive biomarker and its mechanism in the progression of liver cirrhosis.Methods:Gene Expression Omnibus(GEO)dataset(GSE15654,n=216)was analyzed to screen genes associated with progression of liver cirrhosis.A total of 181 plasma samples,including healthy control(HC,n=20),chronic hepatitis B(CHB,n=77)and HBV-related liver cirrhosis(LC,n=84),were enrolled for validation.In vitro and in vivo experiments were employed for the mechanistic investigation.Results:GEO dataset analysis showed that relatively low mRNA-expression of C–C motif chemokine ligand 16(CCL16)was associated with elevated Child-Pugh score(P=0.034)and worse prognosis(P=0.025).Plasma CCL16 level decreased in a stepwise pattern,with a median concentration of 10.29,6.57 and 4.47 ng/mL in the HC,CHB and LC groups,respectively(P<0.001).Low plasma CCL16 was significantly related to hepatic dysfunction both in the CHB and LC groups(P<0.05).Combination of CCL16 and ALT showed improved distinguishing capability for LC compared to either alone.In vitro,CCL16 expression was downregulated by lipopolysaccharide and hypoxia.Overexpression of CCL16 from human normal liver cell line(LO2)reduced the extracellular matrix associated proteins(Col1 and Col4)in human hepatic stellate cell line(LX-2).In vivo,the pathological feature of cirrhosis was alleviated by the hepatocytespecific expression of CCL16.Conclusions:CCL16 could be a feasible plasma marker to predict the occurrence and progression of liver cirrhosis.CCL16 might impact liver cirrhosis through inactivating hepatic stellate cells.

CCL16 inhibits tumor proliferation and metastasis in HCC by impacting CK19 phenotype

Background and aims:Cytokeratin 19-positive(CK19t)hepatocellular carcinoma(HCC)is an aggressive subtype with poor outcomes.The initiation and development of CK19t HCC in the background of liver cirrhosis remains unclear.This study investigated the role of the cirrhosis-related gene C-C motif chemokine ligand 16(CCL16)in the development of CK19t HCC.Methods:Datasets from Gene Expression Omnibus(GEO),The Cancer Genome Atlas(TCGA)and International Cancer Genome Consortium(ICGC)were analyzed to screen and validate the genes associated with CK19t HCC.A total of 102 HCC patients were included for tissue microarray analysis.Gain-of-function experiments were conducted to investigate the biological functions of CCL16.CIBERSORT was used to investigate the correlation of CCL16 and immune infiltration.Results:GEO dataset analysis showed that CK19t HCC had lower expression of CCL16.In both TCGA dataset and our HCC cohort,CCL16 expression was negatively correlated with CK19 expression(P<0.05)and its expression was higher in para-tumor than tumor tissues(P<0.001).Moreover,low CCL16 expression was related to advanced stage and poor overall survival(P<0.05).CCL16 overexpression downregulated CK19 expression and impacted the sphere formation ability of HCC cells.Overexpression of CCL16 inhibited the cell proliferation,migration,and invasion of HCC cell lines.Immune analysis showed HCC with high CCL16 expression had more infiltration of mast cells.HCC patients with both low CCL16 expression and low mast cells had the worst prognosis(P<0.001).Conclusion:Our data indicated that CCL16 downregulated the expression of CK19 and inhibited the malignant phenotype of HCC.