Gastroenterology

Gastroenterology

Volume 125, Issue 1, July 2003, Pages 89-97
Gastroenterology

Clinical-liver, pancreas, and biliary tract
Glypican-3: a novel serum and histochemical marker for hepatocellular carcinoma

https://doi.org/10.1016/S0016-5085(03)00689-9Get rights and content

Abstract

Background & aims:

Early detection of hepatocellular carcinoma (HCC) is critical for successful treatment. However, the differential diagnosis between HCC and benign hepatic lesions is sometimes difficult and new biochemical markers for HCC are required. It has been reported that glypican-3 (GPC3) messenger RNA (mRNA) is significantly increased in most HCCs compared with benign liver lesions or normal liver. The goal of this study is to determine whether GPC3 is also overexpressed at the protein level and whether GPC3 is detectable in the serum of patients with HCC.

Methods:

GPC3 was assessed in liver tissue sections by immunohistochemistry and in serum by enzyme-linked immunosorbent assay. Serum α-fetoprotein (AFP) level was also measured in the same patients.

Results:

Immunohistochemical studies showed that GPC3 is expressed in 72% of HCCs (21 of 29), whereas it is not detectable in hepatocytes from normal liver and benign liver diseases. Consistent with this, GPC3 was undetectable in the serum of healthy donors and patients with hepatitis, but its levels were significantly increased in 18 of 34 patients (53%) with HCC. In addition, only 1 of 20 patients with hepatitis plus liver cirrhosis displayed elevated levels of serum GPC3. Interestingly, in most cases, there was no correlation between GPC3 and AFP values. Thus, at least 1 of the 2 markers was elevated in 82% of the patients with HCC.

Conclusions:

GPC3 is specifically overexpressed in most HCCs and is elevated in the serum of a large proportion of patients with HCC. The simultaneous determination of GPC3 and AFP may significantly increase the sensitivity for diagnosis of HCC.

Section snippets

Patient tissues

Liver tissue samples were obtained from the Department of Pathology at Toronto General Hospital. All liver specimens were large blocks from surgically resected tumors and adjacent parenchyma. All tissue was fixed in 10% formalin and embedded in paraffin for routine histologic examination.

Patient sera

Blood samples were obtained from 34 patients with HCC (see Table 1 for patient characteristics), 20 patients with hepatitis plus liver cirrhosis (12 with hepatitis C virus and 8 with hepatitis B virus; 17 with

Generation of anti-GPC3 mouse mAbs

To generate the antibodies, we used a His-tagged carboxyl-terminal GPC3 fragment as immunogen. After screening the hybridomas by ELISA against the immunogen without the His tag, 13 positive clones were selected. The positive clones were then tested by immunostaining GPC3-transfected 293 cells. Two stable hybridoma clones, named 1G12 and 8H5, immunostained the GPC3-expressing cells in a strong and specific manner (Figure 1). Because both antibodies displayed similar reactivity, we chose the

Discussion

By immunostaining paraffin-embedded tissue sections with an mAb, we have shown that GPC3 is expressed in 72% of HCCs, whereas it is undetectable in hepatocytes from benign liver diseases, reactive liver, and normal liver. These results are consistent with the 2 previous studies that assessed the levels of GPC3 mRNA in HCC and nonmalignant hepatic tissue18, 19 and suggest that GPC3 could be a useful immunohistochemical marker to distinguish between benign and malignant hepatocellular mass

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    M.C. is supported by a fellowship from the Cancer Research Society of Canada.

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