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

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Abstract

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.

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Data availability

The authors declare that all the data supporting the findings of this study are available within the article and its supplementary information files.

Abbreviations

GPC3:

Glypican3

GS:

Glutamine synthetase

HCC:

Hepatocellular carcinoma

IHC:

Immunohistochemistry

ICCs:

Intrahepatic cholangiocarcinomas

HBV:

Hepatitis B virus

WHO:

World Health Organization

HCV:

Hepatitis C virus

ANOVA:

Analysis of variance

SEM:

Standard error of mean

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Acknowledgements

We gratefully acknowledge the support from the National Natural Science Foundation of China (No. 81572309), the Natural Science Foundation of Guangdong Province (No. 2017A030313502 and No. 2018A030313650 and No. 2019A1515011455), and the Guangzhou Science and Technology Project (No. 201707010119), Guangdong Province, China.

Funding

This work was supported by the National Natural Science Foundation of China (No. 81572309), the Natural Science Foundation of Guangdong Province (No. 2017A030313502 and No. 2018A030313650 and No. 2019A1515011455), and the Guangzhou Science and Technology Project (No. 201707010119), Guangdong Province, China. The authors thank the Department of Pathology of The Third Affiliated Hospital of Sun Yat-sen University.

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Authors and Affiliations

  1. Department of Pathology, The Third Affiliated Hospital, Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, People’s Republic of China

    Ahmed Musa Hago Bakheet, Chang Zhao, Jian-Ning Chen, Jing-Yue Zhang, Jun-Ting Huang, Yu Du, Li-Ping Gong, Yuan-Hua Bi & Chun-Kui Shao

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  1. Ahmed Musa Hago Bakheet
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  2. Chang Zhao
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Contributions

CKS contributed to study concept, design and supervised all works; AMHB contributed to the experiments, analysis, and interpretation of data and drafting of the manuscript. CZ, JNC contributed to samples’ pathological diagnosis and staging. JYZ contributed to acquisition of data. JTH, YD, LPG, and YHB contributed to technical and material support.

Corresponding author

Correspondence to Chun-Kui Shao.

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Conflict of interest

Ahmed Musa Hago Bakheet, Chang Zhao, Jian-Ning Chen, Jing-Yue Zhang, Jun-Ting Huang, Yu Du, Li-Ping Gong, Yuan-Hua Bi, and Chun-Kui Shao declare that no conflicts of interest exist. The authors have no financial relationship to disclose.

Ethical approval

We declare that the authors used the archived wax-embedded blocks of liver samples in the Pathology Department at the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. This article did not show patients’ personal information and the medical ethics involved in the research were examined and approved by the Ethics Committee of Sun Yat-sen University, Guangzhou, China. None of the patients treated by chemotherapy or radiotherapy before the operation and the medical ethics involved in the research were examined and approved by the Ethics Committee of Sun Yat-sen University, Guangzhou, China.

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Chang Zhao is the co-first author.

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Bakheet, A.M.H., Zhao, C., Chen, JN. et al. Improving pathological early diagnosis and differential biomarker value for hepatocellular carcinoma via RNAscope technology. Hepatol Int 14, 96–104 (2020). https://doi.org/10.1007/s12072-019-10006-z

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