Skip to main content

Advertisement

Springer Nature Link
Log in

CDK4/6 inhibitors downregulate the ubiquitin-conjugating enzymes UBE2C/S/T involved in the ubiquitin–proteasome pathway in ER + breast cancer

  • Research Article
  • Published:
Clinical and Translational Oncology Aims and scope Submit manuscript

A Correction to this article was published on 10 September 2022

This article has been updated

Abstract

Purpose

Despite significant improvement in therapeutic development in the past decades, breast cancer remains a formidable cause of death for women worldwide. The hormone positive subtype (HR(+)) (also known as luminal type) is the most prevalant category of breast cancer, comprising ~70% of patients. The clinical success of the three CDK4/6 inhibitors palbociclib, ribociclib, and abemaciclib has revolutionized the treatment of choice for metastatic HR(+) breast cancer. Accumulating evidence demonstrate that the properties of CDK4/6 inhibitors extend beyond inhibition of the cell cycle, including modulation of immune function, sensitizing PI3K inhibitors, metabolism reprogramming, kinome rewiring, modulation of the proteosome, and many others. The ubiquitin–proteasome pathway (UPP) is a crucial cellular proteolytic system that maintains the homeostasis and turnover of proteins.

Methods

We performed transcriptional profiling of the HR(+) breast cancer cell lines MCF7 and T47D treated with Palbociclib. Differential expressed genes were analyzed for novel pathways enriched. The results were further validated with biochemical assays and with real world clinical database cohorts.

Results

We uncovered a novel mechanism that demonstrate the CDK4/6 inhibitors suppress the expression of three ubiquitin conjugating enzymes UBE2C, UBE2S, UBE2T. Further validation in the HR(+) cell lines show that Palbociclib and ribociclib decrease UBE2C at both the mRNA and protein level, but this phenomenon was not shared with abemaciclib. These three E2 enzymes modulate several E3 ubiquitin ligases, including the APC/C complex which plays a role in G1/S progression. We further demonstrate the UBE2C/UBE2T expression levels are associated with breast cancer survival, and HR(+) breast cancer cells demonstrate dependence on the UBE2C.

Conclusions

Our study suggests a novel link between CDK4/6 inhibitor and UPP pathway, adding to the potential mechanisms of their clinical efficacy in cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Change history

References

  1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality Worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021;71(3):209–49.

    PubMed  Google Scholar 

  2. Li Y, Yang D, Yin X, Zhang X, Huang J, Wu Y, et al. Clinicopathological characteristics and breast cancer-specific survival of patients with single hormone receptor-positive breast cancer. JAMA Netw Open. 2020;3(1):e1918160.

    PubMed  PubMed Central  Google Scholar 

  3. Chi D, Singhal H, Li L, Xiao T, Liu W, Pun M, et al. Estrogen receptor signaling is reprogrammed during breast tumorigenesis. Proc Natl Acad Sci USA. 2019;116(23):11437–43.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Cardoso F, Paluch-Shimon S, Senkus E, Curigliano G, Aapro MS, Andre F, et al. 5th ESO-ESMO International consensus guidelines for advanced breast cancer (ABC 5). Ann Oncol. 2020;31(12):1623–49.

    CAS  PubMed  Google Scholar 

  5. Finn RS, Martin M, Rugo HS, Jones S, Im SA, Gelmon K, et al. Palbociclib and Letrozole in advanced breast cancer. N Engl J Med. 2016;375(20):1925–36.

    CAS  PubMed  Google Scholar 

  6. Hortobagyi GN, Stemmer SM, Burris HA, Yap YS, Sonke GS, Paluch-Shimon S, et al. Ribociclib as first-line therapy for HR-positive, advanced breast cancer. N Engl J Med. 2016;375(18):1738–48.

    CAS  PubMed  Google Scholar 

  7. Goetz MP, Toi M, Campone M, Sohn J, Paluch-Shimon S, Huober J, et al. MONARCH 3: Abemaciclib as initial therapy for advanced breast cancer. J Clin Oncol off J Am Soc Clin Oncol. 2017;35(32):3638–46.

    CAS  Google Scholar 

  8. Knudsen ES, Shapiro GI, Keyomarsi K. Selective CDK4/6 inhibitors: biologic outcomes, determinants of sensitivity, mechanisms of resistance, combinatorial approaches, and pharmacodynamic biomarkers. Am Soc Clin Oncol Educ Book. 2020;40:115–26.

    PubMed  PubMed Central  Google Scholar 

  9. Goel S, DeCristo MJ, McAllister SS, Zhao JJ. CDK4/6 Inhibition in cancer: beyond cell cycle arrest. Trends Cell Biol. 2018;28(11):911–25.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Goel S, DeCristo MJ, Watt AC, BrinJones H, Sceneay J, Li BB, et al. CDK4/6 inhibition triggers anti-tumour immunity. Nature. 2017;548(7668):471–5.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Lelliott EJ, Kong IY, Zethoven M, Ramsbottom KM, Martelotto LG, Meyran D, et al. CDK4/6 inhibition promotes antitumor immunity through the induction of t-cell memory. Cancer Discov. 2021;11(10):2582–601.

    CAS  PubMed  Google Scholar 

  12. Vora SR, Juric D, Kim N, Mino-Kenudson M, Huynh T, Costa C, et al. CDK 4/6 inhibitors sensitize PIK3CA mutant breast cancer to PI3K inhibitors. Cancer Cell. 2014;26(1):136–49.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Franco J, Balaji U, Freinkman E, Witkiewicz AK, Knudsen ES. Metabolic reprogramming of pancreatic cancer mediated by CDK4/6 inhibition elicits unique vulnerabilities. Cell Rep. 2016;14(5):979–90.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Pancholi S, Ribas R, Simigdala N, Schuster E, Nikitorowicz-Buniak J, Ressa A, et al. Tumour kinome re-wiring governs resistance to palbociclib in oestrogen receptor positive breast cancers, highlighting new therapeutic modalities. Oncogene. 2020;39(25):4781–97.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Klein ME, Kovatcheva M, Davis LE, Tap WD, Koff A. CDK4/6 inhibitors: the mechanism of action may not be as simple as once thought. Cancer Cell. 2018;34(1):9–20.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Du Q, Guo X, Wang M, Li Y, Sun X, Li Q. The application and prospect of CDK4/6 inhibitors in malignant solid tumors. J Hematol Oncol. 2020;13(1):41.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Pernas S, Tolaney SM, Winer EP, Goel S. CDK4/6 inhibition in breast cancer: current practice and future directions. Ther Adv Med Oncol. 2018;10:1758835918786451.

    PubMed  PubMed Central  Google Scholar 

  18. Scheffner M, Nuber U, Huibregtse JM. Protein ubiquitination involving an E1-E2-E3 enzyme ubiquitin thioester cascade. Nature. 1995;373(6509):81–3.

    CAS  PubMed  Google Scholar 

  19. Pickart CM, Eddins MJ. Ubiquitin: structures, functions, mechanisms. Biochem Biophys Acta. 2004;1695(1–3):55–72.

    CAS  PubMed  Google Scholar 

  20. George AJ, Hoffiz YC, Charles AJ, Zhu Y, Mabb AM. A Comprehensive atlas of E3 Ubiquitin ligase mutations in neurological disorders. Front Genet. 2018;9:29.

    PubMed  PubMed Central  Google Scholar 

  21. Miettinen TP, Peltier J, Hartlova A, Gierlinski M, Jansen VM, Trost M, et al. Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib. EMBO J. 2018;37(10):e98359.

    PubMed  PubMed Central  Google Scholar 

  22. Kariri Y, Toss MS, Alsaleem M, Elsharawy KA, Joseph C, Mongan NP, et al. Ubiquitin-conjugating enzyme 2C (UBE2C) is a poor prognostic biomarker in invasive breast cancer. Breast Cancer Res Treat. 2022;192(3):529–39.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Shan BQ, Wang XM, Zheng L, Han Y, Gao J, Lv MD, et al. DCAF13 promotes breast cancer cell proliferation by ubiquitin inhibiting PERP expression. Cancer Sci. 2022;113(5):1589.

    Google Scholar 

  24. Huang W, Liu X, Zhang Y, Deng M, Li G, Chen G, et al. USP5 promotes breast cancer cell proliferation and metastasis by stabilizing HIF2alpha. J Cell Physiol. 2022;237(4):2211–9.

    CAS  PubMed  Google Scholar 

  25. Howley BV, Mohanty B, Dalton A, Grelet S, Karam J, Dincman T, et al. The ubiquitin E3 ligase ARIH1 regulates hnRNP E1 protein stability EMT and breast cancer progression. Oncogene. 2022;41(12):1679–90.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Martinez-Chacin RC, Bodrug T, Bolhuis DL, Kedziora KM, Bonacci T, Ordureau A, et al. Ubiquitin chain-elongating enzyme UBE2S activates the RING E3 ligase APC/C for substrate priming. Nat Struct Mol Biol. 2020;27(6):550–60.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Liess AKL, Kucerova A, Schweimer K, Schlesinger D, Dybkov O, Urlaub H, et al. Dimerization regulates the human APC/C-associated ubiquitin-conjugating enzyme UBE2S. Sci Signal. 2020. https://doi.org/10.1126/scisignal.aba8208.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Kernan J, Bonacci T, Emanuele MJ. Who guards the guardian? mechanisms that restrain APC/C during the cell cycle. Biochim Biophys Acta Mol Cell Res. 2018;1865(12):1924–33.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Hu D, Qiao X, Wu G, Wan Y. The emerging role of APC/CCdh1 in development. Semin Cell Dev Biol. 2011;22(6):579–85.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. da Huang W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 2009;4(1):44–57.

    CAS  Google Scholar 

  31. Huang DW, Sherman BT, Tan Q, Kir J, Liu D, Bryant D, et al. DAVID bioinformatics resources: expanded annotation database and novel algorithms to better extract biology from large gene lists. Nucleic Acids Res. 2007;35(2):W169–75.

    PubMed  PubMed Central  Google Scholar 

  32. Lanczky A, Gyorffy B. Web-based survival analysis tool tailored for medical research (KMplot): development and implementation. J Med Internet Res. 2021;23(7):e27633.

    PubMed  PubMed Central  Google Scholar 

  33. Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z. GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res. 2017;45(W1):W98-102.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Tsherniak A, Vazquez F, Montgomery PG, Weir BA, Kryukov G, Cowley GS, et al. Defining a cancer dependency map. Cell. 2017;170(3):564-76.e16.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Wagner V, Gil J. Senescence as a therapeutically relevant response to CDK4/6 inhibitors. Oncogene. 2020;39(29):5165–76.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Crozier L, Foy R, Mouery BL, Whitaker RH, Corno A, Spanos C, et al. CDK4/6 inhibitors induce replication stress to cause long-term cell cycle withdrawal. EMBO J. 2022;41(6):e108599.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Zhang Z, Li J, Ou Y, Yang G, Deng K, Wang Q, et al. CDK4/6 inhibition blocks cancer metastasis through a USP51-ZEB1-dependent deubiquitination mechanism. Signal Transduct Target Ther. 2020;5(1):25.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Kuner R, Falth M, Pressinotti NC, Brase JC, Puig SB, Metzger J, et al. The maternal embryonic leucine zipper kinase (MELK) is upregulated in high-grade prostate cancer. J Mol Med (Berl). 2013;91(2):237–48.

    CAS  Google Scholar 

  39. O’Leary B, Finn RS, Turner NC. Treating cancer with selective CDK4/6 inhibitors. Nat Rev Clin Oncol. 2016;13(7):417–30.

    CAS  PubMed  Google Scholar 

  40. Harbeck N, Rastogi P, Martin M, Tolaney SM, Shao ZM, Fasching PA, et al. Adjuvant abemaciclib combined with endocrine therapy for high-risk early breast cancer: updated efficacy and Ki-67 analysis from the monarchE study. Ann Oncol. 2021;32(12):1571–81.

    CAS  PubMed  Google Scholar 

  41. Pandey K, An HJ, Kim SK, Lee SA, Kim S, Lim SM, et al. Molecular mechanisms of resistance to CDK4/6 inhibitors in breast cancer: a review. Int J Cancer. 2019;145(5):1179–88.

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Wander SA, Cohen O, Gong X, Johnson GN, Buendia-Buendia JE, Lloyd MR, et al. The genomic landscape of intrinsic and acquired resistance to cyclin-dependent kinase 4/6 inhibitors in patients with hormone receptor-positive metastatic breast cancer. Cancer Discov. 2020;10(8):1174–93.

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Yeon Hee Park S-AI, Kyunghee Park, Ji Wen, Ahrum Min, Vinicius Bonato, Seri Park, Sripad Ram, Dae-Won Lee, Ji-Yeon Kim, Kyung-Hun Lee, Won-Chul Lee, Jisook Lee, Hyunseon(Ally) Kim, Won-Woo Lee, Yoon-La Choi, Scott Weinrich, Han Suk Ryu, Woong-Yang Park, Zhengyan Kan, Palbociclib Biomarker Translational Research Group (Palbo-Bio-TRG). Prospective longitudinal multi-omics study of palbociclib resistance in hormone receptor+/HER2- metastatic breast cancer. ASCO Annu Meet. 2021;39(15):1013–23.

    Google Scholar 

  44. Klein FG, Granier C, Zhao Y, Pan Q, Tong Z, Gschwend JE, et al. Combination of Talazoparib and Palbociclib as a potent treatment strategy in bladder cancer. J Pers Med. 2021;11(5):340.

    PubMed  PubMed Central  Google Scholar 

  45. Li S, Zhang Y, Wang N, Guo R, Liu Q, Lv C, et al. Pan-cancer analysis reveals synergistic effects of CDK4/6i and PARPi combination treatment in RB-proficient and RB-deficient breast cancer cells. Cell Death Dis. 2020;11(4):219.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Brinkmann K, Schell M, Hoppe T, Kashkar H. Regulation of the DNA damage response by ubiquitin conjugation. Front Genet. 2015;6:98.

    PubMed  PubMed Central  Google Scholar 

  47. Jacquemont C, Taniguchi T. Proteasome function is required for DNA damage response and fanconi anemia pathway activation. Can Res. 2007;67(15):7395–405.

    CAS  Google Scholar 

  48. Marra A, Curigliano G. Are all cyclin-dependent kinases 4/6 inhibitors created equal? NPJ Breast Cancer. 2019;5:27.

    PubMed  PubMed Central  Google Scholar 

  49. Pham HT, Nguyen TTT, Nguyen LP, Han SS, Lim YS, Hwang SB. Hepatitis C virus downregulates ubiquitin-conjugating enzyme e2s expression to prevent proteasomal degradation of NS5A, leading to host cells more sensitive to DNA damage. J Virol. 2019;93(2):e01240-e1318.

    PubMed  PubMed Central  Google Scholar 

  50. Sacaan AI, Thibault S, Hong M, Kondegowda NG, Nichols T, Li R, et al. CDK4/6 inhibition on glucose and pancreatic beta cell homeostasis in young and aged rats. Mol Cancer Res. 2017;15(11):1531–41.

    CAS  PubMed  Google Scholar 

  51. Liu Y, Zhao R, Chi S, Zhang W, Xiao C, Zhou X, et al. UBE2C Is upregulated by estrogen and promotes epithelial-mesenchymal transition via p53 in endometrial cancer. Mol Cancer Res. 2020;18(2):204–15.

    CAS  PubMed  Google Scholar 

  52. Min M, Mevissen TE, De Luca M, Komander D, Lindon C. Efficient APC/C substrate degradation in cells undergoing mitotic exit depends on K11 ubiquitin linkages. Mol Biol Cell. 2015;26(24):4325–32.

    CAS  PubMed  PubMed Central  Google Scholar 

  53. Watson ER, Brown NG, Peters JM, Stark H, Schulman BA. Posing the APC/C E3 ubiquitin ligase to orchestrate cell division. Trends Cell Biol. 2019;29(2):117–34.

    CAS  PubMed  Google Scholar 

  54. Mizrak A, Morgan DO. Polyanions provide selective control of APC/C interactions with the activator subunit. Nat Commun. 2019;10(1):5807.

    CAS  PubMed  PubMed Central  Google Scholar 

  55. Binne UK, Classon MK, Dick FA, Wei W, Rape M, Kaelin WG Jr, et al. Retinoblastoma protein and anaphase-promoting complex physically interact and functionally cooperate during cell-cycle exit. Nat Cell Biol. 2007;9(2):225–32.

    CAS  PubMed  Google Scholar 

  56. Mo CH, Gao L, Zhu XF, Wei KL, Zeng JJ, Chen G, et al. The clinicopathological significance of UBE2C in breast cancer: a study based on immunohistochemistry, microarray and RNA-sequencing data. Cancer Cell Int. 2017;17:83.

    PubMed  PubMed Central  Google Scholar 

  57. Psyrri A, Kalogeras KT, Kronenwett R, Wirtz RM, Batistatou A, Bournakis E, et al. Prognostic significance of UBE2C mRNA expression in high-risk early breast cancer a Hellenic Cooperative Oncology Group (HeCOG) Study. Ann Oncol. 2012;23(6):1422–7.

    CAS  PubMed  Google Scholar 

  58. Bavi P, Uddin S, Ahmed M, Jehan Z, Bu R, Abubaker J, et al. Bortezomib stabilizes mitotic cyclins and prevents cell cycle progression via inhibition of UBE2C in colorectal carcinoma. Am J Pathol. 2011;178(5):2109–20.

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Wang H, Zhang C, Rorick A, Wu D, Chiu M, Thomas-Ahner J, et al. CCI-779 inhibits cell-cycle G2-M progression and invasion of castration-resistant prostate cancer via attenuation of UBE2C transcription and mRNA stability. Can Res. 2011;71(14):4866–76.

    CAS  Google Scholar 

  60. Kim YJ, Lee G, Han J, Song K, Choi JS, Choi YL, et al. UBE2C overexpression aggravates patient outcome by promoting estrogen-dependent/independent cell proliferation in early hormone receptor-positive and HER2-negative breast cancer. Front Oncol. 2019;9:1574.

    PubMed  Google Scholar 

Download references

Acknowledgements

This study was partially funded by the grants to JIL: MOST 110-2314-B-A49A-549 (Ministry of Science and Technology), MLCF-V111_A11102 (Melissa Lee Cancer Foundation), Taiwan Clinical Oncology Research Foundation, and 109DHA0100490 (Taipei Veterans General Hospital internal grant).

Author information

Authors and Affiliations

  1. Institute of Clinical Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan

    Chih-Yi Lin, Chung-Jen Yu & Jiun-I. Lai

  2. School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan

    Chun-Yu Liu, Ta-Chung Chao & Ling-Ming Tseng

  3. Division of Medical Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan

    Chun-Yu Liu, Ta-Chung Chao & Jiun-I. Lai

  4. Division of Transfusion Medicine, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan

    Chun-Yu Liu

  5. Comprehensive Breast Health Center, Taipei Veterans General Hospital, Taipei, Taiwan

    Chun-Yu Liu, Ta-Chung Chao, Chi-Cheng Huang, Ling-Ming Tseng & Jiun-I. Lai

  6. Division of General Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan

    Chi-Cheng Huang & Ling-Ming Tseng

  7. Center of Immuno-Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan

    Jiun-I. Lai

Authors
  1. Chih-Yi Lin
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Chung-Jen Yu
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Chun-Yu Liu
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Ta-Chung Chao
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Chi-Cheng Huang
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Ling-Ming Tseng
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Jiun-I. Lai
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence to Jiun-I. Lai.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

For this type of study formal consent is not required.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The original online version of this article was revised to structure the Abstract and to correct the formatting of the Results section.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 57 KB)

Rights and permissions

Springer Nature or its licensor holds exclusive rights to this article under a publishing agreementwith the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lin, CY., Yu, CJ., Liu, CY. et al. CDK4/6 inhibitors downregulate the ubiquitin-conjugating enzymes UBE2C/S/T involved in the ubiquitin–proteasome pathway in ER + breast cancer. Clin Transl Oncol 24, 2120–2135 (2022). https://doi.org/10.1007/s12094-022-02881-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12094-022-02881-0

Keywords