Abstract
Alcohol-related hepatocellular carcinoma (HCC) develops with advanced alcoholic liver disease and liver fibrosis. Using adult mice, we evaluate the effect of alcoholic steatohepatitis on early hepatobiliary carcinoma after initiation by diethyl-nitrosamine (DEN). Here we show that alcohol-fed DEN-injected mice have higher ALT and liver-to-body weight ratio compared to pair-fed DEN-injected mice. Alcohol feeding results in steatohepatitis indicated by increased pro-inflammatory cytokines and fibrotic genes. MRI and liver histology of alcohol+DEN mice shows hepatobiliary cysts, early hepatic neoplasia and increase in serum alpha-fetoprotein. Proliferation makers (BrdU, cyclin D1, p53) and cancer stem cell markers (CD133 and nanog) are significantly up-regulated in livers of alcohol-fed DEN-injected mice compared to controls. In livers with tumors, loss of miR-122 expression with a significant up-regulation of miR-122 target HIF-1α is seen. We conclude that alcoholic steatohepatitis accelerates hepatobiliary tumors with characteristic molecular features of HCC by up-regulating inflammation, cell proliferation, stemness, and miR-122 loss.
Highlights
By thiobarbituric acid reactive substances (TBARS) assay. (e) Representative Sirius Red staining images from all experimental groups
We found that the expression of miR-122 in the liver tissue was significantly lower in alcohol-fed DEN-injected mice compared to any other groups in this study (Fig. 6a). miR-122 regulates the expression of cyclin G1, whose high levels have been reported in several human cancers[30]
In this study we demonstrate that chronic alcohol feeding in adult mice accelerated DEN induced liver tumor development with molecular characteristics of Hepatocellular carcinoma (HCC)
Summary
By TBARS assay. (e) Representative Sirius Red staining images from all experimental groups. (e) Representative Sirius Red staining images from all experimental groups. Bars inside the images indicate 100 μ m. (f) For quantification, at least 3 different microscope fields at 10x magnification were scored for each mouse (n ≥ 5mice per group). Bar graph shows percent Sirius red positive area quantified using ImageJ. (g) Fold changes in mRNA levels of fibrosis markers in liver tissue. Values are given as average ± SD, ANOVA and Dunnett’s multiple comparison were used to compare the means of multiple groups; (*p < 0.05)
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