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METTL3 facilitates stemness properties and tumorigenicity of cancer stem cells in hepatocellular carcinoma through the SOCS3/JAK2/STAT3 signaling pathway

Cancer Gene Therapy volume 31, pages 228–236 (2024)Cite this article

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Abstract

Liver cancer stem cells (LCSCs) contribute to tumor recurrence and cancer cell proliferation in patients with hepatocellular carcinoma (HCC). METTL3-catalyzed m6A modification is relevant to the cancer stem cell (CSC) phenotype, including LCSCs. LCSCs were isolated from MHCC-97H and HepG2 cells through flow cytometry. UALCAN data were used to analyze the expression of METTL3 in liver hepatocellular carcinoma (LIHC) tissues. Loss- and gain-of-function experiments were utilized to assess the biological effects of METTL3 and SOCS3 on the proliferation and stemness phenotypes in vitro and in vivo. The mechanisms underlying the impact of METTL3 were explored using qPCR, MeRIP-qPCR, dual-luciferase reporter, and western blot assays. METTL3 was significantly upregulated in LIHC tissues according to the UALCAN database. METTL3 was highly expressed in LIHC and was significantly correlated with individual cancer stage, tumor grade and lymph node metastasis. Patients with low METTL3 expression had a longer overall survival time based on the data from UALCAN. In addition, the level of METTL3 was enhanced in LCSCs and decreased in non-LCSCs compared to HCC cells. Moreover, overexpression of METTL3 stimulated the proliferation and stemness of LCSCs in vitro and in vivo, while loss of METTL3 impeded it. Bioinformatics analysis combined with validation experiments determined that m6A was modified by METTL3-targeting SOCS3 mRNA. METTL3 had side effects regarding the stability of SOCS3 mRNA. SOCS3 overexpression impaired and SOCS3 depletion facilitated the development of LCSCs via the JAK2/STAT3 pathway. Furthermore, METTL3 depletion suppressed proliferation and stemness in LCSCs, which was restored by SOCS3 knockdown or colivelin treatment. We discovered that METTL3 facilitated the stemness and tumorigenicity of LCSCs by modifying SOCS3 mRNA with m6A.

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Fig. 1: METTL3 could be a potential biomarker indicating poor prognosis of HCC.
Fig. 2: METTL3 was highly expressed in LCSCs.
Fig. 3: METTL3 accelerated the proliferation and stemness of LCSCs.
Fig. 4: METTL3 accelerated the tumorigenicity of LCSCs in vivo.
Fig. 5: METTL3 regulated SOCS3 mRNA stability.
Fig. 6: SOCS3 regulated the proliferation and stemness of LCSCs through the JAK2/STAT3 pathway.
Fig. 7: METTL3-mediated LCSC proliferation and stemness through the SOCS3/JAK2/STAT3 axis.
Fig. 8: Regulatory mechanism of METTL3 in the self-renewal and proliferation of LCSCs.

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Data availability

The datasets used or analyzed during this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We would like to thank the anonymous reviewers who have helped to improve the paper.

Funding

This work was supported by the Ningbo Natural Science Foundation (Grant No.2022J248), Key Research Foundation of Ningbo No. 2 Hospital, China (Grant No. 2022HMZD08), the Public Welfare Foundation of Ningbo (2021S108), Ningbo leading Medical & Health Discipline-Anorectal Surgery (2022-B11), and the Science and Research Project of Ningbo City College of Vocational Technology (ZZX22230).

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Authors and Affiliations

  1. Department of Ultrasonography, Ningbo No. 2 Hospital, 315010, Ningbo, Zhejiang Province, P. R. China

    Jingjing Hu, Fangfang Hong & Hua Yin

  2. Ningbo City College of Vocational Technology, 315100, Ningbo, Zhejiang Province, P. R. China

    Ke Chen

  3. Clinical Laboratory, Ningbo No. 2 Hospital, 315010, Ningbo, Zhejiang Province, P. R. China

    Guosheng Gao

  4. Department of Anorectal Surgery, Ningbo No. 2 Hospital, 315010, Ningbo, Zhejiang Province, P. R. China

    Xiaoyu Dai

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HY guaranteed the integrity of the entire study; JH designed the study and literature research; HY defined the intellectual content; KC and FH performed the experiments; GG collected the data; XD analyzed the data; and HY wrote the main manuscript and prepared the figures. All authors reviewed the manuscript.

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Correspondence to Hua Yin.

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This study was approved by the ethics committee of Ningbo No. 2 Hospital. This article does not contain any studies with human participants performed by any of the authors.

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Hu, J., Chen, K., Hong, F. et al. METTL3 facilitates stemness properties and tumorigenicity of cancer stem cells in hepatocellular carcinoma through the SOCS3/JAK2/STAT3 signaling pathway. Cancer Gene Ther 31, 228–236 (2024). https://doi.org/10.1038/s41417-023-00697-w

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