Diagnostic Value of Glypican3, Heat Shock Protein 70 and Glutamine Synthetase in Hepatocellular Carcinoma Arising in Cirrhotic and Non-Cirrhotic Livers

Abstract

Background and objectives: Histopathological distinction of various nodular lesions in liver with sufficient sensitivity and specificity is a challenge even in an expert set up. The panel of immunohistochemical markers composed of glutamine synthetase (GS), Glypican3 (GPC3) and heat shock protein 70 (HSP70) was recommended by the International Consensus Group for Hepatocellular Neoplasia group for the differentiation of high grade dysplastic nodule and early hepatocellular carcinoma (HCC). The panel has been extensively validated in the western population. This study aims to test this panel on Indian population on resected, explanted and autopsy cirrhotic and non-cirrhotic liver specimens of HCC.

Methodology: This study was conducted on 39 such liver specimens (12 cirrhotic, 12 pre-cirrhotic and 11 non-cirrhotic, non-fibrotic livers), including 35 cases of HCC over a period of 12 years. Immunohistochemistry was performed with antibodies against GS, GPC3 and HSP70 on the sections containing both malignant and dysplastic nodules.

Results: The diagnostic yield depended upon the nature of background liver pathology and was found to be high for only those HCCs arising in cirrhotic background, when positivity of any two markers was taken to be in favor of HCC (sensitivity-58.33%; specificity-100%). GS had a sensitivity and Negative predictive value of 100% for HCCs arising in cirrhotic livers.

Conclusions: Strong positivity for GS is a highly sensitive marker for HCC in a cirrhotic background regardless of the differentiation of the tumor in Indian population. This may be due to preferential activation of Wnt pathway in Indian patients with cirrhosis. The sensitivity of the panel was too low for detecting HCCs arising in non-cirrhotic livers, even in the pre-cirrhotic chronically inflamed livers, even though the specificity was high. GPC3 and HSP70 appear to be useful as individual markers for HCCs arising in non-cirrhotic livers.

Keywords: C, cirrhotic; GPC3, Glypican3; GS, glutamine synthetase; Glypican3; HCC, hepatocellular carcinoma; HGDN, high grade dysplastic nodule; HSP70, heat shock protein 70; ICGHN, International Consensus Group for Hepatocellular Neoplasia; LGDN, low grade dysplastic nodules; NCNF, non-cirrhotic, non-fibrotic livers; PC, pre-cirrhotic; dysplastic nodules; glutamine synthetase; heat shock protein 70; hepatocellular carcinoma.

Figure 1

(A) Strong cytoplasmic staining of glutamine synthetase (magnification 400×). (B) Differential positivity of glutamine synthetase within the same tumor, adjacent liver parenchyma shows positivity in the perivenular hepatocytes (magnification 40×). (C) Diffuse and strong cytoplasmic of Glypican3 staining in the tumor cells. The adjacent liver parenchyma is negative. (magnification 100×). (D) Strong membranous positivity of Glypican3 in more than 10% tumor cells (magnification 400×).

Figure 2

(A) Strong nucleocytoplasmic staining of heat shock protein 70 (HSP70) in the tumor cells (magnification 400×). (B) Cytoplasmic positivity with no nuclear positivity, considered to be negative (magnification 100×).