Expression of Glypican-3 in Mouse Embryo Stem Cells and its Derived Hepatic Lineage Cells Treated with Diethylnitrosamine in vitro

Abstract

To clarify the role of stem cells in hepatocarcinogenesis, glypican-3 (GPC-3) and E-cadherin expression was investigated in embryonic cell lineages. Mouse embryonic stem cells (ESCs), hepatic progenitor cells (HPCs) and hepatocyte like cells (HCs), representing 0, 22 and 40 days of differentiation, respectively, were treated in vitro with diethylnitrosamine (DEN) at four doses (0, 1, 5 and 15 mM; G1, G2, G3 and G4, respectively) for 24 h and GPC-3 and E-cadherin expression was examined by relative quantitative real-time PCR and immunocytochemistry. GPC-3 mRNA expression was significantly different for G4 at day 0 (p<0.001) and for G4 at day 22 (p<0.01) compared with the control (G1). E-cadherin mRNA expression was significantly different for G3 and G4 at day 0 (p<0.05 and p<0.001, respectively), for G2 and G4 (p<0.05 and p<0.001, respectively) at day 22 and for G2 and G4 (p<0.01 and p<0.001, respectively) at day 40 compared with G1. Immunofluorescence staining for GPC-3 showed a membranous and/or granular expression in cytoplasm of ESCs and HPCs and granular and/or diffuse expression in cytoplasm of HCs, which were also stained by E-cadherin. DEN treatment increased GPC-3 expression in ESCs, HPCs and HCs, with increase of E-cadherin expression. Taken together, the expression of GPC-3 was altered by DEN treatment. However, its expression pattern was different at the stage of embryo stem cells and its derived hepatic lineage cells. This suggests that GPC-3 expression may be modulated in the progeny of stem cells during their differentiation toward hepatocytes, associated with E-cadherin expression.

Differential expression of glypican-3 (GPC3) in lung squamous cell carcinoma and lung adenocarcinoma and its clinical significance

“…This protein is distributed on the cell surface and in the extracellular matrix. Mutation and abnormal expression of GPC3 protein have been associated with malignant tumors (Umezu et al, 2010;Kim and Kang, 2013;Greten et al, 2013;Feng and Ho, 2014). GPC3 is a member of the superfamily of membrane-bound heparan sulfate proteoglycans.…”

The significance of glypican‐3 expression profiling in the tumor cellular origin theoretical system for hepatocellular carcinoma progression

“…During liver development and differentiation of hepatic parenchymal cells, the presence and withdrawal of GPC3 are better programmed than AFP. The presence of GPC3 and AFP proteins can be traced to the hepatoblast or hepatic stem cells at the same embryonic weeks . With the differentiation and maturation, expressions of both fetal proteins tend to decrease.…”

“…Differentiation of cancer cells had a clinical significance and was found to be positively correlated with the invasive proclivity . GPC3 may be modulated in the progeny of stem cells during their differentiation toward hepatocytes . However, the dynamic expression of GPC3 and its differentiation status for identifying the tumor cellular origins are still obscure.…”

Prediction of Early Recurrence of Solitary Hepatocellular Carcinoma after Orthotopic Liver Transplantation

“…In addition to cholangiocellular carcinoma, CK19 can be detected in some HCC subtypes, indicating that the CK19-positive HCC can originate from HPC 17,18 . GPC3 was abundantly expressed in HPCs and immature hepatocytes but not in mature hepatocytes 19–21 . In well differentiated HCC, GPC3 expression was weak or absent 22,23 .…”