In our earlier work, we demonstrated that agrin, a multifunctional heparan sulfate proteoglycan, accumulates in hepatocellular carcinoma (HCC) and cholangiocellular carcinoma (CCC). In addition, we proved the utility of agrin immumohistochemistry in discriminating between HCCs and benign parenchymal lesions. Here, we have examined the expression of agrin in metastatic liver carcinomas in comparison with primary liver tumors. Immunohistochemistry for agrin was performed on 25 HCC, 16 intrahepatic CCC, 20 colorectal cancer metastasis (CRCm), and 18 pancreatic ductal carcinoma metastasis (PDCm) samples and evaluated with both quantitative and qualitative methods. Agrin/CD34 double immunofluorescent staining was carried out on snap-frozen sections. Agrin mRNA expression was measured in 11 HCC, 7 CCC, 11 CRCm, and 12 normal liver tissues. Regardless of tumor grade, agrin immunostaining was strong in the microvessels of HCCs. As opposed to HCC, agrin immunostaining was faint or nearly absent from the CD34-positive microvessels of CCC, CRCm, and PDCm; rather, it was detected in the basement membranes surrounding tumor cell pseudoglandules. While agrin was preserved in the basement membranes of Grade III CCCs, it was nearly absent from poorly differentiated metastatic adenocarcinomas. Agrin mRNA levels were the highest in CCC and lower, but still elevated in HCC and CRCm. By qualitative evaluation of agrin immunoreactions, CCC was differentiated from CRCm and PDCm with a sensitivity of 0.81 and a specificity of 0.82. HCCs were unequivocally identified on the basis of microvascular agrin labeling. Thus, agrin immunohistochemistry may facilitate determination of primary versus metastatic origin in problematic liver cancer cases.
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