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CSK-homologous kinase (CHK/MATK) is a potential colorectal cancer tumour suppressor gene epigenetically silenced by promoter methylation

Oncogene volume 40, pages 3015–3029 (2021)Cite this article

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Abstract

Hyperactivation of SRC-family protein kinases (SFKs) contributes to the initiation and progression of human colorectal cancer (CRC). Since oncogenic mutations of SFK genes are rare in human CRC, we investigated if SFK hyperactivation is linked to dysregulation of their upstream inhibitors, C-terminal SRC kinase (CSK) and its homolog CSK-homologous kinase (CHK/MATK). We demonstrate that expression of CHK/MATK but not CSK was significantly downregulated in CRC cell lines and primary tumours compared to normal colonic tissue. Investigation of the mechanism by which CHK/MATK expression is down-regulated in CRC cells uncovered hypermethylation of the CHK/MATK promoter in CRC cell lines and primary tumours. Promoter methylation of CHK/MATK was also observed in several other tumour types. Consistent with epigenetic silencing of CHK/MATK, genetic deletion or pharmacological inhibition of DNA methyltransferases increased CHK/MATK mRNA expression in CHK/MATK-methylated colon cancer cell lines. SFKs were hyperactivated in CHK/MATK-methylated CRC cells despite expressing enzymatically active CSK, suggesting loss of CHK/MATK contributes to SFK hyperactivation. Re-expression of CHK/MATK in CRC cell lines led to reduction in SFK activity via a non-catalytic mechanism, a reduction in anchorage-independent growth, cell proliferation and migration in vitro, and a reduction in tumour growth and metastasis in a zebrafish embryo xenotransplantation model in vivo, collectively identifying CHK/MATK as a novel putative tumour suppressor gene in CRC. Furthermore, our discovery that CHK/MATK hypermethylation occurs in the majority of tumours warrants its further investigation as a diagnostic marker of CRC.

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Fig. 1: CHK/MATK expression is suppressed in colorectal tumours.
Fig. 2: Down-regulation of CHK/MATK expression correlates with activation of Src in colorectal cancer cell lines.
Fig. 3: The CHK/MATK promoter is heavily methylated in four representative colorectal cancer cell lines.
Fig. 4: Quantitative MS-PCR analysis showing CHK/MATK promoter hypermethylation in a panel of CRC cell lines and increased CHK/MATK promoter methylation in primary colorectal cancers.
Fig. 5: CHK/MATK hypermethylation in colorectal cancers occurs primarily at the gene promoter and flanking regions.
Fig. 6: Treatment of colorectal cancer cells with the demethylating agent 5-Aza-2′-deoxycytidine (Aza-dC) induces de-repression of CHK/MATK expression.
Fig. 7: Effects of expression of recombinant CHK/MATK on Src kinase activity and phosphorylation state of HCT116 CRC cells.
Fig. 8: Effects of expression of recombinant CHK/MATK in HCT116 CRC cells on cell migration, anchorage-independent growth and proliferation in vitro and on growth and survival in vivo in zebrafish xenograft.

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Funding

O. Sieber is a NHMRC Senior Research Fellow (GNT1136119); grant support to H.-J. Zhu: NHMRC project grant #628727, grant support to H.-C. Cheng: NHMRC project grant # 1050486 and Australian Brain Foundation, grant support to A. Dhillon: NHMRC project grants #1141906. J. Mariadason was supported by a NHMRC Senior Research fellowship (GNT1046092). F. Hollande received grants from the Tour de Cure Foundation, Australia (Senior Project grant) and the National Health and Medical Research Council of Australia (Grant #1164081).

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Authors and Affiliations

  1. Olivia Newton-John Cancer Research Institute, Austin Hospital, Heidelberg, Victoria, Australia

    Anderly C. Chüeh, Ian Luk & John M. Mariadason

  2. The Walter and Eliza Hall Institute, Parkville, Victoria, Australia

    Anderly C. Chüeh, Anuratha Sakthianandeswaren, Dmitri Mouradov & Oliver Sieber

  3. Department of Medical Biology, University of Melbourne, Parkville, Victoria, Australia

    Anderly C. Chüeh, Anuratha Sakthianandeswaren, Dmitri Mouradov & Oliver Sieber

  4. Department of Medicine (Austin Hospital), University of Melbourne, Heidelberg, Victoria, Australia

    Anderly C. Chüeh & John M. Mariadason

  5. Cancer Program and Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia

    Anderly C. Chüeh & Oliver Sieber

  6. Department of Biochemistry and Pharmacology, Bio21 Institute, University of Melbourne, Parkville, Victoria, Australia

    Gahana Advani, Nadia Ng, Daisy S. Lio, Heather Verkade & Heung-Chin Cheng

  7. Department of Clinical Pathology, the University of Melbourne, Victorian Comprehensive Cancer Centre, Parkville, Victoria, Australia

    Momeneh Foroutan, Jai Smith & Frédéric Hollande

  8. The University of Melbourne Centre for Cancer Research, Victorian Comprehensive Cancer Centre, Parkville, Victoria, Australia

    Momeneh Foroutan, Jai Smith & Frédéric Hollande

  9. Australian Centre for Blood Diseases, Monash University, Prahran, Victoria, Australia

    Harshal Nandurkar

  10. Department of Surgery (Royal Melbourne Hospital), University of Melbourne, Parkville, Victoria, Australia

    Hong-Jian Zhu & Oliver Sieber

  11. Discipline of Laboratory Medicine, School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria, Australia

    Yuh-Ping Chong

  12. Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada

    Donald J. Fujita & Jeffrey Bjorge

  13. The Institute for Mental and Physical Health and Clinical Translation, School of Medicine, Deakin University, Geelong, Victoria, Australia

    Faiza Basheer, Jet Phey Lim & Amardeep Dhillon

  14. School of Cancer Medicine, La Trobe University, Heidelberg, Victoria, Australia

    John M. Mariadason

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  1. Anderly C. Chüeh
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Contributions

Conception, experimental design and data analysis: ACC, GA, NN, H-CC, H-JZ, HV, HN, JB, DJF, DM, OS, FH, AD, FB and JM. Performing experiments: ACC, GA, NN, MF, JS, DSL, H-JZ, JB, IL, AS and FB. Preparation of the manuscript: ACC, Y-PC, JPL, GA, NN, H-CC, H-JZ, HV, HN, JB, DJF, OS, FH, FB, AD and JM.

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Correspondence to Anderly C. Chüeh or Heung-Chin Cheng.

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Chüeh, A.C., Advani, G., Foroutan, M. et al. CSK-homologous kinase (CHK/MATK) is a potential colorectal cancer tumour suppressor gene epigenetically silenced by promoter methylation. Oncogene 40, 3015–3029 (2021). https://doi.org/10.1038/s41388-021-01755-z

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