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Sirt1 deficiency upregulates glutathione metabolism to prevent hepatocellular carcinoma initiation in mice

Oncogene volume 40, pages 6023–6033 (2021)Cite this article

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Abstract

Sirtuin-1 (SIRT1) is involved in various metabolic pathways, including fatty acid synthesis and gluconeogenesis in the liver. However, its role in initiation and progression of liver cancer remains unclear. Studying Sirt1 liver-specific knockout (LKO) mice in combination with diethylnitrosamine (DEN) treatment, we demonstrated that loss of Sirt1 rendered mice resistant to DEN-induced hepatocellular carcinoma (HCC) development. RNA-seq revealed that livers from LKO mice exhibited an enrichment in glutathione metabolism eight months after DEN challenge. Sirt1 deficiency elevated the expression of glutathione-s-transferase family genes by increasing the level of Nrf2, a key regulator of glutathione metabolism. Hence, LKO livers displayed a reductive environment with an increased ratio of GSH to GSSG and an elevated GSH level. Furthermore, using CRISPR knockout techniques, we confirmed that the impairment of HCC formation in LKO mice is mainly dependent on NRF2 signaling. Meanwhile, HCC induced by DEN could be blocked by the administration of N-acetyl cysteine (NAC) when administered one month after DEN challenge. However, NAC treatment starting five months after DEN injection was not able to prevent tumor development. In conclusion, our findings indicate that a reductive environment orchestrated by glutathione metabolism at an early stage can prevent the initiation of HCC.

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Fig. 1: Sirt1 LKO mice resist HCC formation after DEN challenge.
Fig. 2: Sirt1 deficiency reduces cell proliferation after DEN treatment.
Fig. 3: Sirt1 deficiency enhances glutathione metabolism in liver with DEN treatment.
Fig. 4: Increased Nrf2 activity in LKO regulates glutathione genes in DEN treatment.
Fig. 5: Knockout Nrf2 in Sirt1 LKO mice augments DEN induced HCC growth.
Fig. 6: Pretreatment of antioxidant reduces DEN induced tumor growth.

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Data availability

All RNA-sequencing data generated in this study have been deposited in GEO (https://www.ncbi.nlm.nih.gov/geo/) under the accession number PRJNA728801.

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Acknowledgements

We thank Ms. Ragini Bhalchandra Adhav for helping analyze ChIP-seq data, and other members of Xu lab for helpful discussion. We also thank Dr. Yiwei Cao, Dr. Hao Xiao and Mr. Haibin Yang for collecting animal tissues, and FHS Animal Research Core for providing animal housing.

Funding

This project was supported by grants SRG2015-00008-FHS, MYRG2016-00054-FHS and MYRG2017-00096-FHS to RHW; MYRG2019-0064-FHS to XLX; and CPG2020-00004-FHS to CXD from the University of Macau.

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Authors and Affiliations

  1. Cancer Center, Faculty of Health Sciences, University of Macau, Macau, SAR, China

    Pengxiang Qiu, Weilong Hou, Haitao Wang, Kimmy Ka Wing Lei, Shaowei Wang, Lakhansing Arun Pardeshi, Samson Sek Man Su, Qiang Chen, Chu-Xia Deng, Xiaoling Xu & Ruihong Wang

  2. Center for Precision Medicine Research and Training, Faculty of Health Sciences, University of Macau, Macau, SAR, China

    Pengxiang Qiu, Samson Sek Man Su, Qiang Chen, Chu-Xia Deng & Xiaoling Xu

  3. National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, USA

    Weiping Chen

  4. Thoracic Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Katherine Prothro, Yashvita Shukla, David S. Schrump & Ruihong Wang

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Contributions

PQ, WH: designed and optimized experiments, analyzed and interpreted data, drafted and edited the final paper; PQ, WH, HW, WC: analysis of data, preparing figures; KKWL, SW, SSMS, LAP, and QC: optimized experiments; KP, YS, and DSS: edited and reviewed final paper; CXD, XX, and RW: designed the project, analyzed and interpreted the data, edited and finalized the paper.

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Correspondence to Chu-Xia Deng, Xiaoling Xu or Ruihong Wang.

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Qiu, P., Hou, W., Wang, H. et al. Sirt1 deficiency upregulates glutathione metabolism to prevent hepatocellular carcinoma initiation in mice. Oncogene 40, 6023–6033 (2021). https://doi.org/10.1038/s41388-021-01993-1

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