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Ubiquitination-specific protease 7 enhances stemness of hepatocellular carcinoma by stabilizing basic transcription factor 3

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Abstract

This study aims to explore the molecular regulation mechanism of ubiquitination-specific protease 7 (USP7) in facilitating the stemness properties of hepatocellular carcinoma (HCC). Gain-of-function and loss-of-function assays were conducted in SK-Hep1 and HepG2 cells transfected with USP7 overexpression/knockdown plasmids and USP7 inhibitor P22077. The proliferation, migration, invasion, and self-renewal capacity of hepatocellular carcinoma cells were detected by CCK-8, colony formation, Transwell, scratch, and tumor sphere formation, respectively. MS was performed to identify the potential substrate of USP7 following P22077 treatment. Co-IP assay was used to verify the interaction between USP7 and basic transcription factor 3 (BTF3) in HCC cells. The overexpression of USP7 could promote the proliferation, migration, invasion, and colony formation capacity of SK-Hep1 and HepG2 cells. Additionally, ectopic UPS7 enhanced the epithelial-mesenchymal transition (EMT) and stem-like characteristics of the HCC cells. In contrast, USP7 depletion by knockdown of USP7 or administrating inhibitor P22077 significantly inhibited these malignant phenotypes of SK-Hep1 and HepG2 cells. Following MS analysis, BTF3 was identified as a potential substrate for USP7. USP7 could interact with BTF3 and upregulate its protein level, while USP7 depletion significantly upregulated the ubiquitination levels. Overexpression of BTF3 partially rescue the inhibitory effects of USP7 depletion on the malignant phenotypes and stemness properties of SK-Hep1 and HepG2 cells. USP7 can promote the stemness and malignant phenotype of HCC by stabilizing BTF3.

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Funding

This study was supported by grants from National Natural Science Foundation (82272839, 81702419) and Postdoctoral Science Foundation of Jiangsu Province (2021K243B).

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Authors and Affiliations

  1. Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Medical School of Nantong University, 20 Xisi Road, Nantong, 226001, Jiangsu, China

    Mingchao Hu, Chengchen Dai, Xieyin Sun, Yinqi Chen, Nuo Xu, Zhaoyi Lin, Shiyu Xu, Saiyan Bian & Wenjie Zheng

  2. Department of Oncology, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong, 226001, China

    Mingchao Hu, Chengchen Dai, Chun Cheng & Wenjie Zheng

  3. Department of Nuclear Medicine, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong, 226001, China

    Mingchao Hu & Zhonghua Tan

Authors
  1. Mingchao Hu
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  2. Chengchen Dai
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  3. Xieyin Sun
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  4. Yinqi Chen
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  5. Nuo Xu
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  6. Zhaoyi Lin
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  7. Shiyu Xu
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  8. Chun Cheng
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  9. Zhonghua Tan
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  10. Saiyan Bian
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  11. Wenjie Zheng
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Contributions

W. Z., Z. T., and C. D. designed and supervised this study. M. H., Y. C., C. D., and S. B. conducted the experiments and drafted the manuscript. N. X., X. S., Z. L., and S. X. analyzed data and performed the statistics. All authors read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Zhonghua Tan, Saiyan Bian or Wenjie Zheng.

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Mingchao Hu, Chengchen Dai, and Xieyin Sun contributed equally to this study.

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Hu, M., Dai, C., Sun, X. et al. Ubiquitination-specific protease 7 enhances stemness of hepatocellular carcinoma by stabilizing basic transcription factor 3. Funct Integr Genomics 24, 28 (2024). https://doi.org/10.1007/s10142-024-01310-5

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