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O-GlcNAcylation of TRIM29 and OGT translation forms a feedback loop to promote adaptive response of PDAC cells to glucose deficiency

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Abstract

Purpose

Glucose not only provides energy for tumor cells, but also provides various biomolecules that are essential for their survival, proliferation and invasion. Therefore, it is of great clinical significance to understand the mechanism of how tumor cells adapt to metabolic stress and maintain their survival. The aim of this research was to study the critical role of OGT and TRIM29 O-GlcNAc modification driven adaptability of PDAC cells to low glucose stress, which might have important medical implications for PDAC therapy.

Methods

Western blotting, mass spectrometry and WGA-immunoprecipitation were used to examined the levels of OGT and O-GlcNAc glycosylated proteins in BxPC3 and SW1990 cells in normal culture and under glucose deprivation conditions. Crystal violet assay, flow cytometry, RIP, RT-qPCR, protein stability assay, biotin pull down were used to investigate the mechanism of OGT and TRIM29-mediated adaptive response to glucose deficiency in PDAC cells.

Results

The current study found that under the condition of low glucose culture, the levels of OGT and O-GlcNAc glycosylation in PDAC cells were significantly higher than those in normal culture. Moreover, the high expression of OGT has a protective effect on PDAC cells under low glucose stress. This study confirmed that there was no significant change in mRNA level and protein degradation of OGT under low glucose stress, which was mainly reflected in the increase of protein synthesis. In addition, O-GlcNAc modification at T120 site plays a critical role in the metabolic adaptive responses mediated by TRIM29.

Conclusions

Taken together, our study indicated that O-GlcNAcylation of TRIM29 at T120 site and OGT translation forms a loop feedback to facilitate survival of PDAC under glucose deficiency.

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Data availability

All data in this study will be available from the corresponding author on reasonable request.

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Acknowledgements

We thank Key Laboratory of Cell Biology in Ministry of Public Health, and Key Laboratory of Medical Cell Biology in Ministry of Education of China Medical University for providing experimental platform.

Funding

This work was partly supported by National Natural Science Foundation of China (32071258, 82203337); Natural Science Foundation of the Higher Education Institutions of Jiangsu Province, China (22KJB320017); Supporting project of Nanjing University of Chinese Medicine to NSFC (XPT82203337); Doctoral Start-up Foundation of Liaoning Province (2021-BS-093); Doctoral Start-up Foundation of Liaoning Province (2020-BS-101).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, China Medical University, Shenyang, 110122, China

    Fu-Ying Zhao, Xue Chen, Chao Li & Hua-Qin Wang

  2. Department of Biochemistry and Molecular Biology, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Jia Sun

  3. Department of Laboratory Medicine, The 1st Affiliated Hospital, China Medical University, Shenyang, 110001, China

    Jia-Mei Wang

  4. Central Laboratory, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, 110042, China

    Ye Yuan

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Contributions

JS and HW designed this study; FZ, XC, JW and YY performed the experiments; XC, JS, JW, CL analyzed the data; HW and FZ wrote the manuscript. All authors have contributed to the final version of the manuscript.

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Correspondence to Jia Sun or Hua-Qin Wang.

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Zhao, FY., Chen, X., Wang, JM. et al. O-GlcNAcylation of TRIM29 and OGT translation forms a feedback loop to promote adaptive response of PDAC cells to glucose deficiency. Cell Oncol. (2024). https://doi.org/10.1007/s13402-023-00915-5

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