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Involvement of CDK11B-mediated SPDEF ubiquitination and SPDEF-mediated microRNA-448 activation in the oncogenicity and self-renewal of hepatocellular carcinoma stem cells

Cancer Gene Therapy volume 28, pages 1136–1149 (2021)Cite this article

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Abstract

Increasing evidence has suggested the crucial role cyclin-dependent kinases (CDKs) in the biology of hepatocellular carcinoma (HCC), a lethal malignancy with high morbidity and mortality. Hence, this study explored the modulatory effect of the putative cyclin-dependent kinase 11B (CDK11B)-mediated ubiquitination on HCC stem cells. The expression of CDK11B, SAM pointed domain-containing ETS transcription factor (SPDEF) and DOT1-like histone lysine methyltransferase (DOT1L) was determined by RT-qPCR and western blot analysis in HCC tissues and cells. The interaction among CDK11B, SPDEF, miR-448, and DOT1L was analyzed by Co-IP, ubiquitination-IP and ChIP assays, whereas their effects on the biological characteristics of HCC stem cells were assessed by sphere formation and colony formation assays. An in vivo xenograft tumor model was developed for validating the regulation of CDK11B in oncogenicity of HCC stem cells. We characterized the aberrant upregulation of CDK11B and downregulation SPDEF in HCC tissues and cells. CDK11B degraded SPDEF through ubiquitin-proteasome pathway, whereas SPDEF could bind to the miR-448 promoter and inhibit the expression of DOT1L by activating miR-448, whereby promoting self-renewal of HCC stem cells. Knockdown of CDK11B attenuated the self-renewal capability of HCC stem cells and their oncogenicity in vivo. These findings highlighted that blocking the CDK11B-induced degradation of SPDEF and enhancing miR-448-dependent inhibition of DOT1L may delay the progression of HCC by restraining self-renewal capability of HCC stem cells, representing novel targets for HCC management.

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Fig. 1: CDK11B is highly expressed in HCC and its knockdown reduces the proliferation of HCC cells.
Fig. 2: CDK11B suppresses the expression of SPDEF by promoting SPDEF phosphorylation and ubiquitination.
Fig. 3: SPDEF inhibits DOT1L expression by activating miR-448 transcription.
Fig. 4: The self-renewal of HCC SMMC-7721 stem cells was inhibited by SPDEF through downregulation of DOT1L repression.
Fig. 5: The involvement of the CDK11B/SPDEF/miR-448/DOT1L axis in the oncogenicity and self-renewal property of HCC stem cells in vivo.
Fig. 6: A schematic for the CDK11B/SPDEF/miR-448/DOT1L axis in the progression of HCC.

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Acknowledgements

We acknowledge and appreciate our colleagues for their valuable suggestions and technical assistance for this study. This work was supported by the Key Research and Development Plan of Hainan Province (2019RC373) and Hainan Natural Science Foundation (818MS161).

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, Central South University Xiangya School of Medicine Affiliated Haikou Hospital, Haikou, 570208, P. R. China

    Jun-cheng Guo, Yi-jun Yang & Jian-quan Zhang

  2. Psychological Research Center, Hainan General Hospital, Haikou, 570311, P. R. China

    Min Guo

  3. Department of Hepatobiliary Surgery, Hainan General Hospital, Haikou, 570311, P. R. China

    Jin-fang Zheng

  4. School of Public Health, Hainan Medical University, Haikou, 571199, P. R. China

    Zhuo Liu

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  1. Jun-cheng Guo
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Jun-cheng Guo and Yi-jun Yang designed the study. Min Guo, Jian-quan Zhang, and Jin-fang Zheng collated the data, carried out data analyses and produced the initial draft of the manuscript. Zhuo Liu contributed to drafting the manuscript. All authors have read and approved the final submitted manuscript.

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Correspondence to Yi-jun Yang or Jian-quan Zhang.

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Guo, Jc., Yang, Yj., Guo, M. et al. Involvement of CDK11B-mediated SPDEF ubiquitination and SPDEF-mediated microRNA-448 activation in the oncogenicity and self-renewal of hepatocellular carcinoma stem cells. Cancer Gene Ther 28, 1136–1149 (2021). https://doi.org/10.1038/s41417-020-00261-w

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