The MyoD family inhibitor domain-containing protein enhances the chemoresistance of cancer stem cells in the epithelial state by increasing β-catenin activity

Abstract

Cancer cells with mesenchymal attributes potentially display chemoresistance. Cancer stem cells (CSCs), which are intrinsically resistant to most chemotherapy agents, exhibit considerable phenotypic heterogeneity in their epithelial versus mesenchymal states. However, the drug response of CSCs in the epithelial and mesenchymal states has not been completely investigated. In this study, we found that epithelial-type (E-cadherinhigh/CD133high) CSCs displayed a higher sphere formation ability and chemoresistance than mesenchymal-type (E-cadherinlowCD133high) CSCs. Gene expression profiling of the CSC and non-CSC subpopulations with distinct epithelial-to-mesenchymal transition (EMT) states showed that MyoD family inhibitor domain-containing (MDFIC) was selectively upregulated in epithelial-type CSCs. Knockdown of MDFIC sensitized epithelial-type CSCs to chemotherapy agents. Ectopic expression of MDFIC increased the chemoresistance of mesenchymal-type CSCs. In a tissue microarray, high MDFIC expression was associated with poor prognosis of non-small cell lung cancer (NSCLC) patients. A mechanistic study showed that the MDFIC p32 isoform, which is located in the cytoplasm, interacted with the destruction complex, Axin/GSK-3/β-catenin. This interaction stabilized β-catenin by inhibiting β-catenin phosphorylation at S33/37 and increased the nuclear translocation and transcriptional activity of β-catenin. Knockdown of β-catenin decreased MDFIC-enhanced chemoresistance. These results suggested that the upregulation of MDFIC enhanced the chemoresistance of epithelial-type CSCs by elevating β-catenin activity. Thus, targeting MDFIC-regulated β-catenin signaling of epithelial-type CSCs may be a potential strategy to overcome chemoresistance in NSCLC.

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Fig. 1: Characterization of the epithelial- and mesenchymal-type CSCs.
Fig. 2: Identification of the drug-resistant signature of the E-cadHCD133H subpopulation.
Fig. 3: MDFIC promotes chemoresistance of the E-cadHCD133H subpopulations of PC14 and A549 cell lines.
Fig. 4: High MDFIC expression correlates with poor overall and disease-free survival of NSCLC patients.
Fig. 5: MDFIC interacts with the Axin/GSK-3/β-catenin complex and stabilizes β-catenin via inhibiting β-catenin phosphorylation at Ser33/37.
Fig. 6: MDFIC contributes to the nuclear translocation and transcriptional activity of β-catenin.

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Acknowledgements

This study was financially supported by the Ministry of Science and Technology of Taiwan (MOST 107-2320-B-037-025-) and Kaohsiung Medical University Hospital (KMUH106-6M63). This work was also financially supported by the Research Center for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan. We thank Dr Michael Hsiao for providing the tissue microarray chips. We thank the assistance of the Department of Pathology and the Biobank in Kaohsiung Medical University Hospital, Kaohsiung, Taiwan for the clinical sample collection. We thank the Center for Research Resources and Development in Kaohsiung Medical University for the instrumental support for the confocal microscope and TissueFAX system. The authors thank the Immunobiology core facility of Clinical Medicine Research Center in National Cheng Kung University Hospital for assisting with the fluorescence-activated cell sorting.

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Funding

This study was financially supported by the Ministry of Science and Technology of Taiwan (MOST 107-2320-B-037-025-) and Kaohsiung Medical University Hospital (KMUH105-5M58).

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Correspondence to Yu-Peng Liu.

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Chen, C., Yang, C., Yang, S. et al. The MyoD family inhibitor domain-containing protein enhances the chemoresistance of cancer stem cells in the epithelial state by increasing β-catenin activity. Oncogene 39, 2377–2390 (2020). https://doi.org/10.1038/s41388-019-1152-4

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