Improving pathological early diagnosis and differential biomarker value for hepatocellular carcinoma via RNAscope technology

Hepatol Int. 2020 Jan;14(1):96-104. doi: 10.1007/s12072-019-10006-z. Epub 2019 Dec 12.


Background: The diagnostic and prognostic values of glypican3 (GPC3) and glutamine synthetase (GS) proteins in hepatocellular carcinoma (HCC) have been reported, but their specificity and sensitivity remain low. Here, we applied RNAscope to improve HCC early pathological and differential diagnosis by estimating GPC3 and GS mRNAs.

Methods: We performed RNAscope and immunohistochemistry (IHC) to detect GPC3 and GS biomarkers on the tissue sections of 194 cases, including high- and low-grade liver dysplastic nodules; highly, moderately, and poorly differentiated HCCs; intrahepatic cholangiocarcinomas (ICCs); metastatic HCC; and carcinomas from other organs.

Results: The results showed that all the cases that were negative for GPC3 by RNAscope were also negative for this protein by IHC. The use of RNAscope assay improved the GPC3 and GS specificity and sensitivity by 20-30%. Hence, HCC shows early recognition and upgrades the metastatic HCC differentiation by 23% compared with IHC (p = 0.0001, 0.0064). Meanwhile, all liver cirrhosis, cholangiocytes and non-HCC samples were negative for GPC3 and GS except lymphocytes in lymphomas, and 2 (8.3%) out of the 24 ICC samples but not in the cancer cells.

Conclusion: RNAscope for GPC3 and GS panel was highly specific and sensitive for the pathological identification of dysplastic nodules, early stages of HCCs, and would differentiate them from HCCs and metastatic tumors compared with IHC.

Keywords: Early diagnosis; Liver cancer; Pathological biomarker; RNAscope.

Publication types

  • Evaluation Study

MeSH terms

  • Biomarkers, Tumor
  • Biopsy / instrumentation*
  • Carcinoma, Hepatocellular / pathology*
  • Early Detection of Cancer
  • Female
  • Glutamate-Ammonia Ligase / genetics*
  • Glypicans / genetics*
  • Humans
  • Immunohistochemistry
  • Liver Neoplasms / pathology*
  • Male
  • Predictive Value of Tests
  • Sensitivity and Specificity


  • Biomarkers, Tumor
  • Glypicans
  • Glutamate-Ammonia Ligase