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DOK7 CpG hypermethylation in blood leukocytes as an epigenetic biomarker for acquired tamoxifen resistant in breast cancer

Journal of Human Genetics volume 68, pages 33–38 (2023)Cite this article

Subjects

Abstract

Background

Breast cancer (BC) is among the most common cause of cancer 10.4% and one of the leading causes of death among 20–50 years old women in the world. Tamoxifen drug is the first line therapy for BC however tamoxifen resistance (TR) has shown in 30–50% of cases that may face BC recurrence. Hence, TR early detection reduces BC recurrence and fatalities. The epigenetic alteration that happens by hypermethylation of tumor suppressor genes and hypomethylation of oncogenes has been suggested to be useful in early cancer or drug resistance diagnosis.

Methods

This is the first study to investigate DOK7 CpG hypermethylation in blood leukocytes of 31 TR (ER+) BC compared to 29 tamoxifen sensitive BC to evaluate DOK7 as a potential TR biomarker. DNA was extracted from blood samples of all participants and MSRE-PCR and real-time PCR were used for quantification of CpG methylation alterations.

Results

The means of DOK7 CpG hypermethylation were obtained as 85.03%, 29.1% and 57.34% in TR, TS and normal control respectively. Significant hypermethylation were found among TR vs. TS (p < 0.001), TS vs. normal (p < 0.001) and TR vs. normal controls (p < 0.03). Online databases expression and survival analysis of DOK7 showed increasing expression in TS groups vs. TR groups which have consistency with our methylation alteration results. The sensitivity and specificity of the TR epigenetic test were determined using ROC analysis showed 89.66% and 96.77% respectively and showed that 37.5% above hypermethylation is at risk for TR and breast cancer recurrence.

Conclusion

There is a significant difference in the methylation ratio of DOK7 between tamoxifen resistant and tamoxifen sensitive groups that may be useful in the early diagnosis of tamoxifen resistance in BC cases and cancer recurrence prevention.

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References

  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. 2020;70:7–30.

    Article  Google Scholar 

  2. Lumachi F, Brunello A, Maruzzo M, Basso U, Basso SM. Treatment of estrogen receptor-positive breast cancer. Curr Med Chem. 2013;20:596–604.

    Article  CAS  Google Scholar 

  3. Tsang JYS, Tse GM. Molecular classification of breast cancer. Adv Anat Pathol. 2020;27:27–35.

    Article  CAS  Google Scholar 

  4. Hon JD, Singh B, Sahin A, Du G, Wang J, Wang VY, et al. Breast cancer molecular subtypes: from TNBC to QNBC. Am J Cancer Res. 2016;6:1864–72.

    CAS  Google Scholar 

  5. Eroles P, Bosch A, Pérez-Fidalgo JA, Lluch A. Molecular biology in breast cancer: intrinsic subtypes and signaling pathways. Cancer Treat Rev. 2012;38:698–707.

    Article  CAS  Google Scholar 

  6. Matthews J, Gustafsson JA. Estrogen signaling: a subtle balance between ER alpha and ER beta. Mol Inter. 2003;3:281–92.

    Article  CAS  Google Scholar 

  7. Hayashi S, Yamaguchi Y. Estrogen signaling pathway and hormonal therapy. Breast Cancer. 2008;15:256–61.

    Article  Google Scholar 

  8. Saha Roy S, Vadlamudi RK. Role of estrogen receptor signaling in breast cancer metastasis. Int J Breast Cancer. 2012;2012:654698.

    Article  Google Scholar 

  9. Levin ER, Pietras RJ. Estrogen receptors outside the nucleus in breast cancer. Breast Cancer Res Treat. 2008;108:351–61.

    Article  CAS  Google Scholar 

  10. Yaşar P, Ayaz G, User SD, Güpür G, Muyan M. Molecular mechanism of estrogen-estrogen receptor signaling. Reprod Med Biol. 2017;16:4–20.

    Article  Google Scholar 

  11. Komm BS, Mirkin S. An overview of current and emerging SERMs. J Steroid Biochem Mol Biol. 2014;143:207–22.

    Article  CAS  Google Scholar 

  12. Hultsch S, Kankainen M, Paavolainen L, Kovanen R-M, Ikonen E, Kangaspeska S, et al. Association of tamoxifen resistance and lipid reprogramming in breast cancer. BMC Cancer. 2018;18:850.

    Article  Google Scholar 

  13. Hu R, Hilakivi-Clarke L, Clarke R. Molecular mechanisms of tamoxifen-associated endometrial cancer (Review). Oncol Lett. 2015;9:1495–501.

    Article  CAS  Google Scholar 

  14. Cook KL, Shajahan AN, Clarke R. Autophagy and endocrine resistance in breast cancer. Expert Rev Anticancer Ther. 2011;11:1283–94.

    Article  CAS  Google Scholar 

  15. Mills JN, Rutkovsky AC, Giordano A. Mechanisms of resistance in estrogen receptor positive breast cancer: overcoming resistance to tamoxifen/aromatase inhibitors. Curr Opin Pharm. 2018;41:59–65.

    Article  CAS  Google Scholar 

  16. Fan W, Chang J, Fu P. Endocrine therapy resistance in breast cancer: current status, possible mechanisms and overcoming strategies. Future Med Chem. 2015;7:1511–9.

    Article  CAS  Google Scholar 

  17. Gutierrez MC, Detre S, Johnston S, Mohsin SK, Shou J, Allred DC, et al. Molecular changes in tamoxifen-resistant breast cancer: relathe tionship between estrogen receptor, HER-2, and p38 mitogen-activated protein kinase. J Clin Oncol. 2005;23:2469–76.

    Article  CAS  Google Scholar 

  18. Li L, Choi JY, Lee KM, Sung H, Park SK, Oze I, et al. DNA methylation in peripheral blood: a potential biomarker for cancer molecular epidemiology. J Epidemiol. 2012;22:384–94.

    Article  Google Scholar 

  19. Rauch TA, Wang Z, Wu X, Kernstine KH, Riggs AD, Pfeifer GP. DNA methylation biomarkers for lung cancer. Tumor Biol. 2012;33:287–96.

    Article  CAS  Google Scholar 

  20. Yi JM. DNA methylation change profiling of colorectal disease: screening towards clinical use. Life. 2021;11:5.

    Article  Google Scholar 

  21. Jones N, Dumont DJ. Recruitment of Dok-R to the EGF receptor through its PTB domain is required for attenuation of Erk MAP kinase activation. Curr Biol. 1999;9:1057–60.

    Article  CAS  Google Scholar 

  22. Bedirian A, Baldwin C, Abe J, Takano T, Lemay S. Pleckstrin homology and phosphotyrosine-binding domain-dependent membrane association and tyrosine phosphorylation of Dok-4, an inhibitory adapter molecule expressed in epithelial cells. J Biol Chem. 2004;279:19335–49.

    Article  CAS  Google Scholar 

  23. Okada K, Inoue A, Okada M, Murata Y, Kakuta S, Jigami T, et al. The muscle protein Dok-7 is essential for neuromuscular synaptogenesis. Science. 2006;312:1802–5.

    Article  CAS  Google Scholar 

  24. Bergamin E, Hallock PT, Burden SJ, Hubbard SR. The cytoplasmic adaptor protein Dok7 activates the receptor tyrosine kinase MuSK via dimerization. Mol Cell. 2010;39:100–9.

    Article  CAS  Google Scholar 

  25. Alsallum MS, Alshareef A, Abuzinadah AR, Bamaga AK, Dallol A. A novel DOK7 mutation causing congenital myasthenic syndrome with limb-girdle weakness: case series of three family members. Heliyon. 2021;7:e06869.

    Article  CAS  Google Scholar 

  26. Zhang L, Li R, Hu K, Dai Y, Pang Y, Jiao Y, et al. Prognostic role of DOK family adapters in acute myeloid leukemia. Cancer Gene Ther. 2019;26:305–12.

    Article  CAS  Google Scholar 

  27. Chen G, Yu H, Satherley L, Zabkiewicz C, Resaul J, Zhao H, et al. The downstream of tyrosine kinase 7 is reduced in lung cancer and is associated with poor survival of patients with lung cancer. Oncol Rep. 2017;37:2695–701.

    Article  CAS  Google Scholar 

  28. Hua C-D, Bian E-B, Chen E-F, Yang Z-H, Tang F, Wang H-L, et al. Repression of Dok7 expression mediated by DNMT1 promotes glioma cells proliferation. Biomed Pharmacother. 2018;106:678–85.

    Article  CAS  Google Scholar 

  29. Yang S-M, Li S-Y, Yu H-B, Li J-R, Sun L-L. Repression of DOK7 mediated by DNMT3A promotes the proliferation and invasion of KYSE410 and TE-12 ESCC cells. Biomed Pharmacother. 2017;90:93–9.

    Article  CAS  Google Scholar 

  30. Yue C, Bai Y, Piao Y, Liu H. DOK7 inhibits cell proliferation, migration, and invasion of breast cancer via the PI3K/PTEN/AKT pathway. J Oncol. 2021;2021:4035257.

    Article  Google Scholar 

  31. Heyn H, Carmona FJ, Gomez A, Ferreira HJ, Bell JT, Sayols S, et al. DNA methylation profiling in breast cancer discordant identical twins identifies DOK7 as novel epigenetic biomarker. Carcinogenesis. 2013;34:102–8.

    Article  CAS  Google Scholar 

  32. Oakes CC, La Salle S, Robaire B, Trasler JM. Evaluation of a quantitative DNA methylation analysis technique using methylation-sensitive/dependent restriction enzymes and real-time PCR. Epigenetics. 2006;1:146–52.

    Article  Google Scholar 

  33. Győrffy B. Survival analysis across the entire transcriptome identifies biomarkers with the highest prognostic power in breast cancer. Comput Struct Biotechnol J. 2021;19:4101–9.

    Article  Google Scholar 

  34. Fekete JT, Győrffy B. ROCplot.org: validating predictive biomarkers of chemotherapy/hormonal therapy/anti-HER2 therapy using transcriptomic data of 3,104 breast cancer patients. Int J Cancer. 2019;145:3140–51.

    Article  CAS  Google Scholar 

  35. Riggins RB, Schrecengost RS, Guerrero MS, Bouton AH. Pathways to tamoxifen resistance. Cancer Lett. 2007;256:1–24.

    Article  CAS  Google Scholar 

  36. Flanagan JM, Munoz-Alegre M, Henderson S, Tang T, Sun P, Johnson N, et al. Gene-body hypermethylation of ATM in peripheral blood DNA of bilateral breast cancer patients. Hum Mol Genet. 2009;18:1332–42.

    Article  CAS  Google Scholar 

  37. Iwamoto T, Yamamoto N, Taguchi T, Tamaki Y, Noguchi S. BRCA1 promoter methylation in peripheral blood cells is associated with increased risk of breast cancer with BRCA1 promoter methylation. Breast Cancer Res Treat. 2011;129:69–77.

    Article  CAS  Google Scholar 

  38. Yin Y, Lei S, Li L, Yang X, Yin Q, Xu T, et al. RPTOR methylation in the peripheral blood and breast cancer in the Chinese population. Genes Genomics. 2022;44:435–43.

    Article  CAS  Google Scholar 

  39. Yang R, Pfütze K, Zucknick M, Sutter C, Wappenschmidt B, Marme F, et al. DNA methylation array analyses identified breast cancer-associated HYAL2 methylation in peripheral blood. Int J Cancer. 2015;136:1845–55.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was carried out at the National Institute of Genetic Engineering and Biotechnology (NIGEB), and the authors would like to express their appreciation to NIGEB. We are grateful to Arash Moradi, a student at the National Institute of Genetic Engineering and Biotechnology (NIGEB) for his participation in the study.

Funding

Our study was supported by grants number 766 from National Institute of Genetic Engineering and Biotechnology (NIGEB).

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Authors and Affiliations

  1. Department of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

    Erfan Gowdini, Seyed Ahmad Aleyasin & Newsha Ramezani

  2. Department of General Surgery, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Nahid Nafisi

  3. Medical Physics Department, Iran University of Medical Sciences, Tehran, Iran

    Mahdieh Tutuni

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  1. Erfan Gowdini
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  2. Seyed Ahmad Aleyasin
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Contributions

SAA designed experiments, support and helped write the manuscript; EG carried out some experiments and write the manuscript; NR carried out some experiments; NN and MT contributed to sample collection and patients data extraction.

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Correspondence to Seyed Ahmad Aleyasin.

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Gowdini, E., Aleyasin, S.A., Ramezani, N. et al. DOK7 CpG hypermethylation in blood leukocytes as an epigenetic biomarker for acquired tamoxifen resistant in breast cancer. J Hum Genet 68, 33–38 (2023). https://doi.org/10.1038/s10038-022-01092-3

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