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USP17L2-SIRT7 axis regulates DNA damage repair and chemoresistance in breast cancer cells

  • Preclinical study
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Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Purpose

Sirtuin7 (SIRT7), as a member of the sirtuin and NAD+-dependent protein-modifying enzyme family, plays an important role in regulating cellular metabolism, stress responses, tumorigenesis, and aging. Ubiquitination and deubiquitination are reversible post-translational modifications that regulate protein stability, enzyme activity, protein–protein interactions, and cellular signaling transduction. However, whether SIRT7 is regulated by deubiquitination signaling is unclear. This study aims to elucidate the molecular mechanism of SIRT7 via deubiquitination signaling.

Methods

USP17L2 or SIRT7-targeting shRNAs were used to deplete USP17L2 or SIRT7. Western blot was applied to assess the effects of USP17L2 or SIRT7 depletion. A co-immunoprecipitation assay was used to detect the interaction relationship. Cell Counting Kit-8 assays were applied to assess the viability of breast cancer cells. An immunohistochemistry assay was employed to detect the protein level in samples from breast cancer patients, and the TCGA database was applied to analyze the survival rate of breast cancer patients. Statistical analyses were performed with the Student’s t test (two-tailed unpaired) and χ2 test.

Results

We find that the deubiquitinase USP17L2 interacts with and deubiquitinates SIRT7, thereby increasing SIRT7 protein stability. In addition, USP17L2 regulates DNA damage repair through SIRT7. Furthermore, SIRT7 polyubiquitination is increased by knocking down of USP17L2, which leads to cancer cells sensitizing to chemotherapy. In breast cancer patient samples, high expression of USP17L2 is correlated with increased levels of SIRT7 protein. In conclusion, our study demonstrates that the USP17L2-SIRT7 axis is the new regulator in DNA damage response and chemo-response, suggesting that USP17L2 may be a prognostic factor and a potential therapeutic target in breast cancer.

Conclusion

Our results highlighted that USP17L2 regulates the chemoresistance of breast cancer cells in a SIRT7-dependent manner. Moreover, the role of USP17L2 as a potential therapeutic target in breast cancer and a prognostic factor for patients was elucidated.

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Data availability

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

Abbreviations

SIRT7:

Sirtuin7

HR:

Homologous recombination repair

NHEJ:

Non-homologous end joining

ATM:

Ataxia-telangiectasia mutated

5-FU:

5-Fluorouracil

CDK1:

Cyclin-dependent kinase 1

DUBs:

Deubiquitinating enzymes

UPS:

Ubiquitin–proteasome system

ATCC:

American Type Culture Collection

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

IHC:

Immunohistochemical

WT:

Wild-type

CA mutant:

Catalytically inactive mutant

Co-IP:

Co-immunoprecipitation

IP:

Immunoprecipitated

CHX:

Cycloheximide

CCK-8:

Cell counting Kit-8

DSBs:

DNA double-strand breaks

HDAC:

Histone deacetylase

AMPK:

Adenosine 5′-monophosphate (AMP)-activated protein kinase

MALAT1:

Metastasis associated in lung adenocarcinoma transcript 1

NRF2:

NF-E2-related factor 2

BRD4:

Bromodomain-containing protein 4

MCL1:

Myeloid cell leukemia 1

BRCA1:

Breast cancer gene 1

BRCA2:

Breast cancer gene 2

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Acknowledgements

Our study was supported by the National Natural Science Foundation of China (91749115, 32090032, 32070713) and the Natural Science Foundation of Jiangxi Province (20181ACB20021).

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Author notes

  1. Yang Su and Chenming Wu have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Arrhythmias of the Ministry of Education of China, Research Center for Translational Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai, 200092, China

    Yang Su, Chenming Wu, Lei Li, Yuping Chen, Ying Lv & Jian Yuan

  2. Department of Biochemistry and Molecular Biology, Tongji University School of Medicine, Shanghai, 200092, China

    Yang Su & Jian Yuan

  3. Shanghai East hospital Ji’An hospital, Ji’an, 343000, China

    Jian Yuan

  4. Shanghai East Hospital, Postgraduate Training Base of Jinzhou Medical University, Shanghai, 200120, China

    Yiming Chang & Xinshu Wang

  5. Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China

    Xuebing Jia

  6. Department of Thoracic Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, 330000, Jiangxi, China

    Bentong Yu

  7. Department of Thoracic Surgery, China-Japan Friendship Hospital Jiangxi Hospital, Nanchang, 330000, Jiangxi, China

    Bentong Yu

Authors
  1. Yang Su
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  2. Chenming Wu
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  9. Bentong Yu
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Contributions

YS carried out the molecular lab work, participated in data analysis, participated in the design of the study, and drafted the manuscript; CW participated in the design of the study and revised the manuscript; LL, YChen, XW, XJ, YChang, and YL provided experimental technology. BYu guided experimental technology. JY conceived of the study, designed the study, coordinated the study, and helped draft the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Bentong Yu or Jian Yuan.

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Su, Y., Wu, C., Chang, Y. et al. USP17L2-SIRT7 axis regulates DNA damage repair and chemoresistance in breast cancer cells. Breast Cancer Res Treat 196, 31–44 (2022). https://doi.org/10.1007/s10549-022-06711-3

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