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miR-575/RIPK4 axis modulates cell cycle progression and proliferation by inactivating the Wnt/β-catenin signaling pathway through inhibiting RUNX1 in colon cancer

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Abstract

Receptor interacting protein serine/threonine kinase 4 (RIPK4) is widely involved in human cancer development. Nevertheless, its role in colon cancer (COAD) has not been elucidated till now. Our research aimed at exploring the function and underlying molecular mechanism of RIPK4 in COAD progression. Through bioinformatic analyses and RT-qPCR, RIPK4 was discovered to be increased in COAD cells and tissues, and its high level predicted poor prognosis. Loss-of-function assays revealed that RIPK4 silencing suppressed COAD cell growth, induced cell cycle arrest, and enhanced cell apoptosis. In vivo experiments also proved that tumor growth was inhibited by silencing of RIPK4. Luciferase reporter assay validated that RIPK4 was targeted and negatively regulated by miR-575. Western blotting demonstrated that Wnt3a, phosphorylated (p)-GSK-3β, and cytoplasmic and nuclear β-catenin protein levels, β-catenin nuclear translocation, and Cyclin D1, CDK4, Cyclin E, and c-Myc protein levels were reduced by RIPK4 knockdown, which however was reversed by treatment with LiCl, the Wnt/β-catenin pathway activator. LiCl also offset the influence of RIPK4 knockdown on COAD cell growth, cell cycle process, and apoptosis. Finally, RIPK4 downregulation reduced RUNX1 level, which was upregulated in COAD and its high level predicted poor prognosis. RIPK4 is positively associated with RUNX1 in COAD. Overexpressing RUNX1 antagonized the suppression of RIPK4 knockdown on RUNX1, Wnt3a, p-GSK-3β, cytoplasmic β-catenin, nuclear β-catenin, Cyclin D1, CDK4, Cyclin E, and c-Myc levels. Collectively, miR-575/RIPK4 axis repressed COAD progression via inactivating the Wnt/β-catenin pathway through downregulating RUNX1.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors appreciate all the participants providing supports for this study.

Funding

Our project was supported by Health commission of Hubei Province scientific Research Project: WJ2019H129; Natural Science Foundation of Hubei Province: 2019CFB640; Research Projects of Biomedical Center of Hubei Cancer Hospital: 2022SWZX, China.

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Authors and Affiliations

  1. Department of Hepatopancreatobiliary Surgery, Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 16 Zhuodaoquan South Road, Hongshan District, Wuhan, 430079, China

    Qun Wang, Weijun Lu & Dongde Wu

  2. Colorectal Cancer Clinical Research Center of Wuhan, Wuhan, 430079, China

    Qun Wang, Weijun Lu & Li Lu

  3. Colorectal Cancer Clinical Research Center of Hubei Province, Wuhan, 430079, China

    Qun Wang

  4. Department of Gastrointestinal Surgery, Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430079, China

    Li Lu

  5. Tianjin Medical University, Tianjin, 300070, China

    Ruopu Wu

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  1. Qun Wang
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Contributions

Qun Wang was the main designer of this study. Qun Wang, Weijun Lu, Li Lu, and Dongde Wu performed the experiments and analyzed the data. Qun Wang, Weijun Lu, Ruopu Wu, and Dongde Wu drafted the manuscript. All authors read and approved the final manuscript.

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Correspondence to Qun Wang or Dongde Wu.

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The study protocol was approved by the Ethics Committee of Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology. The animal studies were authorized by the Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology Animal Care and Use Committee.

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Wang, Q., Lu, W., Lu, L. et al. miR-575/RIPK4 axis modulates cell cycle progression and proliferation by inactivating the Wnt/β-catenin signaling pathway through inhibiting RUNX1 in colon cancer. Mol Cell Biochem (2024). https://doi.org/10.1007/s11010-024-04938-w

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