Expression of proteins upregulated in hepatocellular carcinoma in patients with alcoholic hepatitis (AH) compared to non-alcoholic steatohepatitis (NASH): An immunohistochemical analysis of candidate proteins

Abstract

Both non-alcoholic steatohepatitis (NASH) and alcoholic hepatitis (AH) can lead to cirrhosis and hepatocellular carcinoma. However, the rate of progression to cirrhosis and tumorigenesis in AH is greater than that in NASH. We asked whether there are differences between the two conditions in the expression levels of proteins involved in the pathogenesis of hepatocellular carcinoma. The proteins tested were presented at the 2017 American Association for the Study of Liver Diseases (AASLD) Liver Meeting as overexpressed in hepatocellular carcinoma: KLF4, SCL19A1, FANCG, HRH-1, DNMT1, DNMT3B, TNFR2, DUSP4, EGFR, Integrin α6, HDACII, PDE3A, BCL-XL, and MTCO2. The expression of these proteins was measured in liver biopsy sections from NASH and AH patients using immunohistochemical staining with fluorescent antibodies and then quantifying the fluorescence intensity morphometrically. In AH patients, levels of all tested proteins except HRH-1 were elevated compared to normal patients. In NASH patients, KLF4, SCL19A1, FANCG, HDACII, BCL-XL levels were increased compared to normal controls while HRH-1, DNMT1 and PDE3A levels were decreased. The relative expression of all proteins studied except BCL-XL was significantly higher in AH compared to NASH. In conclusion, proteins involved in hepatocellular cancer development are more highly expressed in AH compared to NASH and normal liver, which corresponds with the higher rate of tumorigenesis in AH patients compared to NASH patients.

Figure 1

Representative images of fluorescence intensity to measure integrin alpha-6 expression in AH liver (A) and normal control (B). A line is drawn through the image to yield a fluorescence intensity graph; the intensity of the ten highest peaks are measured, excluding nuc lear regions which are highlighted by DAPI (not shown). Three areas per slide are measured in this manner.

Figure 2

Level of expression of proteins upregulated in hepatocellular carcinoma in AH, NASH or normal controls. Expression is measured as fluorescence intensity and displayed as mean +/− standard deviation.

Figure 2

Level of expression of proteins upregulated in hepatocellular carcinoma in AH, NASH or normal controls. Expression is measured as fluorescence intensity and displayed as mean +/− standard deviation.

Figure 2

Level of expression of proteins upregulated in hepatocellular carcinoma in AH, NASH or normal controls. Expression is measured as fluorescence intensity and displayed as mean +/− standard deviation.