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SOX4 reversibly induces phenotypic changes by suppressing the epithelial marker genes in human keratinocytes

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Abstract

Background

SOX4 is a transcription factor belonging to the SOX (Sry-related High Mobility Group [HMG] box) family and plays a pivotal role in various biological processes at various stages of life. SOX4 is also expressed in the skin in adults and has been reported to be involved in wound healing, tumor formation, and metastasis.

Methods and results

In this study, we investigated the role of SOX4 in keratinocyte phenotypic changes. We generated a SOX4-overexpressing keratinocyte cell line that expresses SOX4 in a doxycycline (DOX)-inducible manner. DOX treatment induced a change from a paving stone-like morphology to a spindle-like morphology under microscopic observation. Comprehensive gene analysis by RNA sequencing revealed increased expression of genes related to anatomical morphogenesis and cell differentiation as well as decreased expression of genes related to epithelial formation and keratinization, suggesting that SOX4 induced EMT-like phenotype in keratinocytes. Differentially expressed genes (DEGs) obtained by RNA-seq were confirmed using qRT-PCR. DOX-treated TY-1 SOX4 showed a decrease in the epithelial markers (KRT15, KRT13, KRT5, and CLDN1) and an increase in the mesenchymal marker FN1. Protein expression changes by Western blotting also showed a decrease in the epithelial marker proteins keratin 15, keratin 13, and claudin 1, and an increase in the mesenchymal marker fibronectin. Removal of DOX from DOX-treated cells also restored the epithelial and mesenchymal markers altered by SOX4.

Conclusion

Our results indicate that SOX4 reversibly induces an EMT-like phenotype in human keratinocytes via suppression of epithelial marker genes.

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Data availability

The datasets generated during and/or analyzed during the present study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors are grateful to the Cooperative Research Project Program of Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA Center), and the University of Tsukuba for their help with RNA-seq analysis and advice. We thank Tsukuba i-Laboratory for the RNA-seq library preparation and sequencing. Thanks are due to the Oral Medical Research Center of Fukuoka Dental College for allowing us the use of their equipment, including the CFX96 Real-time System, LAS-4000 imaging system, and BZ-9000.

Funding

This study was supported by JSPS KAKENHI (Grant no. JP17K11659).

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Authors and Affiliations

  1. Department of Physiological Science and Molecular Biology, Fukuoka Dental College, 2-15‐1 Tamura, Sawara-ku, Fukuoka, Fukuoka, 814-0193, Japan

    Yoshiyuki Nagaoka, Yukimasa Takeishi, Kana Takeda & Mitsutoki Hatta

  2. Department of Oral Growth and Development, Fukuoka Dental College, 2-15-1 Tamura, Sawara-ku, Fukuoka, Fukuoka, 814-0193, Japan

    Yuki Miyake & Kana Takeda

  3. Department of Morphological Biology, Fukuoka Dental College, 2-15-1 Tamura, Sawara-ku, Fukuoka, Fukuoka, 814-0193, Japan

    Kazuhiko Okamura

  4. Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8577, Japan

    Yuan Yao, Kaori Motomura, Hiroaki Daitoku & Akiyoshi Fukamizu

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  1. Yoshiyuki Nagaoka
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  2. Yukimasa Takeishi
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Contributions

YN and MH conceived and designed the study. YN and YM performed the experiments. YN, KO, YY, KM, HD, AF, and MH analyzed the data. YN and MH wrote the manuscript. YN, YT, KT, and MH confirmed the authenticity of all the raw data. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Mitsutoki Hatta.

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Nagaoka, Y., Takeishi, Y., Miyake, Y. et al. SOX4 reversibly induces phenotypic changes by suppressing the epithelial marker genes in human keratinocytes. Mol Biol Rep 51, 116 (2024). https://doi.org/10.1007/s11033-023-09035-7

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