Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 12: 1557 - 1563

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.12.003

Innovative research team achievement column

Role of SIRT3 SUMOylation deficiency in the proliferation and chemotherapeutic sensitivity of breast cancer cells MCF7

  • Yan-yun HAO ,
  • Si-hui Yü ,
  • Jing LU ,
  • Xiang GU ,
  • Fan ZHANG ,
  • Jin-ke CHENG ,
  • Tian-shi WANG
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  • 1.Department of Biochemistry and Molecular Cell Biology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China
    2.Department of Obstetrics and Gynecology, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 201620, China
    3.Department of Substance Addiction, Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China
    4.Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
    5.Department of Otolaryngology Head and Neck Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
CHENG Jin-ke, E-mail: jkcheng@shsmu.edu.cn. #Co-corresponding authors.
WANG Tian-shi, E-mail: tianshi777@shsmu.edu.cn

Online published: 2021-01-28

Supported by

National Natural Science Foundation of China(81730082);Special Funding Project of Shanghai Jiao Tong University(19X160010017);Innovative Research Team of High-Level Local Universities in Shanghai(SSMU-ZLCX20180102)

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Abstract

Objective

·To investigate the effects of deSUMOylation of sirtuin 3 (SIRT3) on the proliferation and chemotherapeutic drug resistance in breast cancer cells.

Methods

·In order to construct the MFC7-SIRT3KO cell line, the CRISPR-Cas9 plasmid pX330-sgRNA targeting SIRT3 gene was designed. Then, this cell line was infected with SIRT3 K288R (sumoylation modification site mutant), SIRT3 WT (wild type) or Vector retrovirus to establish the SIRT3 SUMOylation site mutation and control cell lines. Cell counting and microscopic observation were used to determine the changes in tumor cell number and morphology under the same incubation condition. To detect the chemotherapeutic drug resistance of SIRT3 K288R and the control cells, these cells were treated with different concentrations of adriamycin (ADM) for 24 h, 48 h and 72 h.

Results

·The MFC7-SIRT3KO cell line was constructed successfully, and then SIRT3 K288R, SIRT3 WT and Vector cell lines were established. The cell growth curves showed that the growth rate of SIRT3 K288R cells was significantly slower than those of SIRT3 WT and Vector cells under the same incubation condition (P=0.000). This phenotype was also observed under the microscope. The diameter of non-adherent breast cancer cell clumps of SIRT3 K288R cells was smaller than those of the control cells. ADM resistance experiment showed that the activity of SIRT3 K288R cells was significantly higher than those of Vector and SIRT3 WT cells treated with different concentrations of ADM (1.25 μg/mL, 2.5 μg/mL, 5 μg/mL and 10 μg/mL) for 24 h, 48 h and 72 h (P<0.05), and the half maximal inhibitory concentration (IC50) of ADM in the SIRT3 K288R cells was higher than those in the control cells.

Conclusion

·The SIRT3 SUMOylation deficiency inhibits the proliferation of breast cancer cells MCF7, while increases the drug resistance to ADM.

Key words： sirtuin 3 (SIRT3); SUMOylation; breast cancer; adriamycin (ADM); drug resistance

Cite this article

Yan-yun HAO , Si-hui Yü , Jing LU , Xiang GU , Fan ZHANG , Jin-ke CHENG , Tian-shi WANG . Role of SIRT3 SUMOylation deficiency in the proliferation and chemotherapeutic sensitivity of breast cancer cells MCF7[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(12) : 1557 -1563 . DOI: 10.3969/j.issn.1674-8115.2021.12.003

References

1 Pavlova NN, Thompson CB. The emerging hallmarks of cancer metabolism[J]. Cell Metab, 2016, 23(1): 27-47.
2 Martinez-Outschoorn UE, Peiris-Pagés M, Pestell RG, et al. Cancer metabolism: a therapeutic perspective[J]. Nat Rev Clin Oncol, 2017, 14(1): 11-31.
3 Leone RD, Powell JD. Metabolism of immune cells in cancer[J]. Nat Rev Cancer, 2020, 20(9): 516-531.
4 Vyas S, Zaganjor E, Haigis MC. Mitochondria and cancer[J]. Cell, 2016, 166(3): 555-566.
5 Wang YP, Sharda A, Xu SN, et al. Malic enzyme 2 connects the Krebs cycle intermediate fumarate to mitochondrial biogenesis[J]. Cell Metab, 2021, 33(5): 1027-1041.e8.
6 Krug K, Jaehnig EJ, Satpathy S, et al. Proteogenomic landscape of breast cancer tumorigenesis and targeted therapy[J]. Cell, 2020, 183(5): 1436-1456.e31.
7 Carrico C, Meyer JG, He W, et al. The mitochondrial acylome emerges: proteomics, regulation by sirtuins, and metabolic and disease implications[J]. Cell Metab, 2018, 27(3): 497-512.
8 Hebert AS, Dittenhafer-Reed KE, Yu W, et al. Calorie restriction and SIRT3 trigger global reprogramming of the mitochondrial protein acetylome[J]. Mol Cell, 2013, 49(1): 186-199.
9 Inuzuka H, Gao D, Finley LW, et al. Acetylation-dependent regulation of Skp2 function[J]. Cell, 2012, 150(1): 179-193.
10 Dong XC, Jing LM, Wang WX, et al. Down-regulation of SIRT3 promotes ovarian carcinoma metastasis[J]. Biochem Biophys Res Commun, 2016, 475(3): 245-250.
11 Torrens-Mas M, Pons DG, Sastre-Serra J, et al. SIRT3 silencing sensitizes breast cancer cells to cytotoxic treatments through an increment in ROS production[J]. J Cell Biochem, 2017, 118(2): 397-406.
12 Bergaggio E, Riganti C, Garaffo G, et al. IDH2 inhibition enhances proteasome inhibitor responsiveness in hematological malignancies[J]. Blood, 2019, 133(2): 156-167.
13 Kim HS, Patel K, Muldoon-Jacobs K, et al. SIRT3 is a mitochondria-localized tumor suppressor required for maintenance of mitochondrial integrity and metabolism during stress[J]. Cancer Cell, 2010, 17(1): 41-52.
14 Li M, Chiang YL, Lyssiotis CA, et al. Non-oncogene addiction to SIRT3 plays a critical role in lymphomagenesis[J]. Cancer Cell, 2019, 35(6): 916-931.e9.
15 Cheng J, Bawa T, Lee P, et al. Role of desumoylation in the development of prostate cancer[J]. Neoplasia, 2006, 8(8): 667-676.
16 He J, Cheng J, Wang T. SUMOylation-mediated response to mitochondrial stress[J]. Int J Mol Sci, 2020, 21(16): 5657.
17 Seeler JS, Dejean A. SUMO and the robustness of cancer[J]. Nat Rev Cancer, 2017, 17(3): 184-197.
18 Wang T, Cao Y, Zheng Q, et al. SENP1-Sirt3 signaling controls mitochondrial protein acetylation and metabolism[J]. Mol Cell, 2019, 75(4): 823-834.e5.
19 He J, Shangguan X, Zhou W, et al. Glucose limitation activates AMPK coupled SENP1-Sirt3 signalling in mitochondria for T cell memory development[J]. Nat Commun, 2021, 12(1): 4371.
20 DeSantis CE, Ma J, Gaudet MM, et al. Breast cancer statistics, 2019[J]. CA Cancer J Clin, 2019, 69(6): 438-451.
21 Harbeck N, Penault-Llorca F, Cortes J, et al. Breast cancer[J]. Nat Rev Dis Primers, 2019, 5(1): 66.
22 Jabbour E, Ravandi F, Kebriaei P, et al. Salvage chemoimmunotherapy with inotuzumab ozogamicin combined with mini-hyper-CVD for patients with relapsed or refractory Philadelphia chromosome-negative acute lymphoblastic leukemia: a phase 2 clinical trial[J]. JAMA Oncol, 2018, 4(2): 230-234.
23 Horwitz S, O'Connor OA, Pro B, et al. Brentuximab vedotin with chemotherapy for CD30-positive peripheral T-cell lymphoma (ECHELON-2): a global, double-blind, randomised, phase 3 trial[J]. Lancet, 2019, 393(10168): 229-240.
24 Cesarman E, Damania B, Krown SE, et al. Kaposi sarcoma[J]. Nat Rev Dis Primers, 2019, 5(1): 9.
25 Basho RK, Gilcrease M, Murthy RK, et al. Targeting the PI3K/AKT/mTOR pathway for the treatment of mesenchymal triple-negative breast cancer: evidence from a phase 1 trial of mTOR inhibition in combination with liposomal doxorubicin and bevacizumab[J]. JAMA Oncol, 2017, 3(4): 509-515.
26 Cai L, Tu J, Song L, et al. Proteome-wide mapping of endogenous SUMOylation sites in mouse testis[J]. Mol Cell Proteomics, 2017, 16(5): 717-727.
27 Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation[J]. Science, 2009, 324(5930): 1029-1033.
28 Guri Y, Colombi M, Dazert E, et al. mTORC2 promotes tumorigenesis via lipid synthesis[J]. Cancer Cell, 2017, 32(6): 807-823.e12.
29 Bogachek MV, Chen Y, Kulak MV, et al. Sumoylation pathway is required to maintain the basal breast cancer subtype[J]. Cancer Cell, 2014, 25(6): 748-761.
30 Kessler JD, Kahle KT, Sun T, et al. A SUMOylation-dependent transcriptional subprogram is required for Myc-driven tumorigenesis[J]. Science, 2012, 335(6066): 348-353.
31 Morris JR, Boutell C, Keppler M, et al. The SUMO modification pathway is involved in the BRCA1 response to genotoxic stress[J]. Nature, 2009, 462(7275): 886-890.
32 Madeddu C, Gramignano G, Floris C, et al. Role of inflammation and oxidative stress in post-menopausal oestrogen-dependent breast cancer[J]. J Cell Mol Med, 2014, 18(12): 2519-2529.
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