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RIOK1 mediates p53 degradation and radioresistance in colorectal cancer through phosphorylation of G3BP2

Abstract

RIO Kinase 1 (RIOK1) is involved in various pathologies, including cancer. However, the role of RIOK1 in radioresistance of colorectal cancer (CRC) remains largely unknown. In this study, we reported that RIOK1 was overexpressed in rectal cancer tissue with weaker tumor regression after neoadjuvant chemoradiotherapy (neoCRT). Moreover, higher RIOK1 expression predicted a poor prognosis in patients with rectal cancer. Blockade of RIOK1 using Toyocamycin, a pharmacological inhibitor of RIOK1, or by knocking down its expression, decreased the resistance of CRC cells to radiotherapy in vitro and in vivo. A mechanistic study revealed that RIOK1 regulates radioresistance by suppressing the p53 signaling pathway. Furthermore, we found that RIOK1 and Ras-GAP SH3 domain binding protein 2 (G3BP2) interact with each other. RIOK1 phosphorylates G3BP2 at Thr226, which increases the activity of G3BP2. RIOK1-mediated phosphorylation of G3BP2 facilitated ubiquitination of p53 by murine double minute 2 protein (MDM2). Altogether, our study revealed the clinical significance of RIOK1 in CRC, and therapies targeting RIOK1 might alleviate the CRC tumor burden in patients.

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Fig. 1: High RIOK1 levels are associated with TRG and poor prognosis in LARC patients treated with neoCRT.
Fig. 2: Knockdown of RIOK1 inhibits resistance of CRC cells to radiotherapy in vivo and in vitro.
Fig. 3: RIOK1 regulates p53 protein stability in CRC cells.
Fig. 4: RIOK1 directly binds to G3BP2 in CRC cells.
Fig. 5: RIOK1-mediated phosphorylation at T226 is required for the function of G3BP2 in regulating p53 stability.
Fig. 6: RIOK1 promotes resistance of CRC cells to radiotherapy via G3BP2.
Fig. 7: Pharmacological inhibitor of RIOK1 promotes sensitivity of CRC cells to radiotherapy.

Data availability

The RNA-seq datasets analyzed for this study are available in the National Center for Biotechnology Information (https://www.ncbi.nlm.nih.gov/sra/). BioProject accession number: PRJNA827462.

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Acknowledgements

This work was supported by China Postdoctoral Science Foundation (No. 2019M663289), Natural Science Foundation of Guangdong Province (Nos. 2019A1515110144 and 2021A1515111014) and National Natural Science Foundation of China (No. 81572725).

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ZH and JH designed the study. YC and SZ performed most of the experiments. KW and LY performed animal experiments and constructed the expression plasmids. QO and JQ collected biological samples, analyzed the data and made the figures. CZ and HD performed Mass Spectrometry Analysis. ZH and SZ participated in writing the manuscript. All authors read and approved the final manuscript.

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Correspondence to Zhenlin Hou.

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The authors declare no competing interests.

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Approvals from the ethical committee of Sun Yat-sen University Cancer Center and prior patient’s consents were previously obtained for the use of these clinical specimens for research purpose. The animal experiments were conducted according to the Animal Study Guidelines of the Ethics Committee of Sun Yat-sen University.

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Chen, Y., Zhou, S., Wan, K. et al. RIOK1 mediates p53 degradation and radioresistance in colorectal cancer through phosphorylation of G3BP2. Oncogene 41, 3433–3444 (2022). https://doi.org/10.1038/s41388-022-02352-4

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