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GPM6A expression is suppressed in hepatocellular carcinoma through miRNA-96 production

Abstract

GPM6A is a glycoprotein in endothelial cells, and its biological function in the development of hepatocellular carcinoma (HCC) is unknown. Through Affymetrix gene expression microarray and bioinformatic analysis, very low GPM6A expression was found in HCC tissue. The present study aims to explore the function and regulatory mechanism of GPM6A in HCC development and progression. Levels of GPM6A expression in HCC specimens from different disorders and various hepatoma cell lines were determined, and its role on cell proliferation was evaluated in hepatoma cells stably overexpressing GPM6A. Modulation of a specific microRNA (miRNA) on its expression and function was evaluated with miRNA mimetic transfection. Herein, it is reported that much lower GPM6A levels were found in HCC tissues than pericancerous liver tissues and correlated to a poor prognosis. GPM6A overexpression inhibited cell proliferation, suppressed colony formation, migration and invasion in two hepatoma cell types. Available evidence does not support that genetic and epigenetic dysregulation contributes significantly to GPM6A inactivation in HCC. Additional findings demonstrated that miR-96-5p acted directly on the 3′-UTR of the GPM6A gene and significantly decreased its mRNA and protein levels. MiR-96-5p transfection promoted proliferation, migration and invasion of SMMC-7721 and MHCC-97H hepatoma cells; whereas the function of oncogenic microRNA-96 was significantly inhibited in GPM6A-overexpressed hepatoma cells. In conclusion, GPM6A expression in HCC is commonly suppressed regardless its base disease types, and its low expression in HCC tissues is most likely attributed to upregulated miR-96-5p. GPM6A may function as a valuable biomarker for HCC progression and prognosis.

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Fig. 1: GPM6A was downregulated in NASH-HCC specimens.
Fig. 2: GPM6A was downregulated in non-NASH-related HCC specimens and human hepatoma cell lines.
Fig. 3: GPM6A suppressed HCC cell proliferation, and migration activity in vitro.
Fig. 4: MiR-96 was identified as an endogenous regulator of GPM6A expression in HCC cells.
Fig. 5: miR-96 promoted proliferation and migration of hepatoma cells.
Fig. 6: miR-96 did not affect proliferation and migration in GPM6A-OE cells.

Data availability

Affymetrix analysis data is accessible at https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE99807.

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Acknowledgements

Part of this work was presented in the American Association of Cancer Research (AACR) Annual Virtual Meeting II, Session title: Mechanisms of Tumor Suppressor Genes, AACR Abstract # 5970, June 24-26, 2020, Philadelphia, PA, USA; and published as an abstract in the Proceedings of AACR Annual Meeting 2020 (DOI: 10.1158/1538-7445). The authors are grateful to senior technologist Ke Qiao in the Technology Platform of Fudan University School of Basic Medical Sciences and Dr. Juan Ye in the Dept. of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Xiang-Nan Yu in the Dept. of Gastroenterology & Hepatology, Zhongshan Hospital of Fudan University, and Dr. Shengyang Ge in the Department of Abdominal Surgery, Huashan Hospital of Fudan University for their technical assistance.

Funding

This work is supported by the National Natural Science Foundation of China (NSFC #81572356, 81871997 and 82170624), the National Key R&D Program of China (#2016YFE0107400) to J.W.

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Z.R.L.: Design and conduct of most experiments, data collection and analysis, paper preparation; G.X., L.Y.Z. and H.C.: Conduct of some experiments, data collection, and analysis. J.M.Z.: providing clinical specimens and patient information. J.W.: Concept development, experimental design and data analysis, overall supervision and funding support, paper preparation, and finalization.

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Correspondence to Jian Wu.

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The authors declare no competing interests.

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The use of human tissue was approved by the Ethic Committee of Fudan University School of Basic Medical Sciences, and followed guidelines of the Helsinki Declaration and the national, municipal, and university regulations.

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Li, ZR., Xu, G., Zhu, LY. et al. GPM6A expression is suppressed in hepatocellular carcinoma through miRNA-96 production. Lab Invest (2022). https://doi.org/10.1038/s41374-022-00818-3

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