Biopolym. Cell. 2024; 40(2):127-135.
Molecular Biomedicine
Doxorubicin affects expression of the ZFP36 and CTTN genes in MCF7 cell line
1, 2Hubiernatorova A. O., 1Kropyvko S. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143
  2. Institute of animal physiology and genetics, CAS
    89, Rumburska str., Libechov, Czech Republic, 27721

Abstract

Aim. The study aims to investigate the effect of doxorubicin treatment of MCF7 cells on the expression of ZFP36, SH3PXD2A, CTTN, SH3PXD2B, WIPF1 and WASL genes coding for TTP, TKS4, TKS5, N-WASP, WIP and CTTN proteins correspondingly. Methods. Cell culture, RT-qPCR, western-blotting and Alamar Blue assay are used in this study. Results. DXR increases both mRNA and protein levels of TTP, which may limit its use as a prognostic marker; it affects the expression of SH3PXD2A, CTTN and WIPF1, but not CTTN and SH3PXD2B. The observed effect is likely an overall response to DNA damage. Conclusion. Doxorubicin significantly affects the expression of ZFP36 and CTTN genes in MCF7 cells, and further investigations are needed to reveal its effects on the cancer cells physiology.
Keywords: doxorubicin, breast cancer, TTP

References

[1] Wilkinson L, Gathani T. Understanding breast cancer as a global health concern. Br J Radiol. 2022; 95(1130):20211033.
[2] Tsang JYS, Tse GM. Molecular Classification of Breast Cancer. Adv Anat Pathol. 2020; 27(1):27-35.
[3] Comşa Ş, Cîmpean AM, Raica M. The Story of MCF-7 Breast Cancer Cell Line: 40 years of Experience in Research. Anticancer Res. 2015; 35(6):3147-54.
[4] Gerstberger S, Jiang Q, Ganesh K. Metastasis. Cell. 2023; 186(8):1564-79.
[5] Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011; 144(5):646-74.
[6] Murphy DA, Courtneidge SA. The 'ins' and 'outs' of podosomes and invadopodia: characteristics, formation and function. Nat Rev Mol Cell Biol. 2011; 12(7):413-26.
[7] Carpenter CL. Actin cytoskeleton and cell signaling. Crit Care Med. 2000; 28(4 Suppl):N94-9.
[8] Saini Y, Chen J, Patial S. The Tristetraprolin Family of RNA-Binding Proteins in Cancer: Progress and Future Prospects. Cancers (Basel). 2020; 12(6):1539.
[9] Fu M, Blackshear PJ. RNA-binding proteins in immune regulation: a focus on CCCH zinc finger proteins. Nat Rev Immunol. 2017; 17(2):130-43.
[10] Roychoudhury S, Kumar A, Bhatkar D, Sharma NK. Molecular avenues in targeted doxorubicin cancer therapy. Future Oncol. 2020; 16(11):687-700.
[11] Sheibani M, Azizi Y, Shayan M, Nezamoleslami S, Eslami F, Farjoo MH, Dehpour AR. Doxorubicin-Induced Cardiotoxicity: An Overview on Pre-clinical Therapeutic Approaches. Cardiovasc Toxicol. 2022; 22(4):292-310.
[12] Sennepin AD, Charpentier S, Normand T, Sarré C, Legrand A, Mollet LM. Multiple reprobing of Western blots after inactivation of peroxidase activity by its substrate, hydrogen peroxide. Anal Biochem. 2009; 393(1):129-31.
[13] Lee SR, Jin H, Kim WT, Kim WJ, Kim SZ, Leem SH, Kim SM. Tristetraprolin activation by resveratrol inhibits the proliferation and metastasis of colorectal cancer cells. Int J Oncol. 2018; 53(3):1269-78.
[14] Kim DJ, Jang JH, Ham SY, Choi SH, Park SS, Jeong SY, Kim BC, Jeon DY, Lee BJ, Ko BK, Park JW, Cho WJ. Doxorubicin inhibits PD-L1 expression by enhancing TTP-mediated decay of PD-L1 mRNA in cancer cells. Biochem Biophys Res Commun. 2020; 522(2):402-7.
[15] Lee JY, Kim HJ, Yoon NA, Lee WH, Min YJ, Ko BK, Lee BJ, Lee A, Cha HJ, Cho WJ, Park JW. Tumor suppressor p53 plays a key role in induction of both tristetraprolin and let-7 in human cancer cells. Nucleic Acids Res. 2013; 41(11):5614-25.
[16] Wei L, Surma M, Gough G, Shi S, Lambert-Cheatham N, Chang J, Shi J. Dissecting the Mechanisms of Doxorubicin and Oxidative Stress-Induced Cytotoxicity: The Involvement of Actin Cytoskeleton and ROCK1. PLoS One. 2015; 10(7):e0131763.
[17] Bober P, Alexovič M, Tomková Z, Kilík R, Sabo J. RHOA and mDia1 Promotes Apoptosis of Breast Cancer Cells Via a High Dose of Doxorubicin Treatment. Open Life Sci. 2019; 14:619-27.
[18] Tchen CR, Brook M, Saklatvala J, Clark AR. The stability of tristetraprolin mRNA is regulated by mitogen-activated protein kinase p38 and by tristetraprolin itself. J Biol Chem. 2004; 279(31):32393-400.
[19] Ross CR, Brennan-Laun SE, Wilson GM. Tristetraprolin: roles in cancer and senescence. Ageing Res Rev. 2012; 11(4):473-84.
[20] Frenay M, Milano G, Renee N, Pons D, Khater R, François E, Thyss A, Namer M. Pharmacokinetics of weekly low dose doxorubicin. Eur J Cancer Clin Oncol. 1989; 25(2):191-5.
[21] Barpe DR, Rosa DD, Froehlich PE. Pharmacokinetic evaluation of doxorubicin plasma levels in normal and overweight patients with breast cancer and simulation of dose adjustment by different indexes of body mass. Eur J Pharm Sci. 2010; 41(3-4):458-63.
[22] Kudlik G, Takács T, Radnai L, Kurilla A, Szeder B, Koprivanacz K, Merő BL, Buday L, Vas V. Advances in Understanding TKS4 and TKS5: Molecular Scaffolds Regulating Cellular Processes from Podosome and Invadopodium Formation to Differentiation and Tissue Homeostasis. Int J Mol Sci. 2020; 21(21):8117.
[23] Ayala I, Baldassarre M, Giacchetti G, Caldieri G, Tetè S, Luini A, Buccione R. Multiple regulatory inputs converge on cortactin to control invadopodia biogenesis and extracellular matrix degradation. J Cell Sci. 2008; 121(Pt 3):369-78.
[24] Fried S, Matalon O, Noy E, Barda-Saad M. WIP: more than a WASp-interacting protein. J Leukoc Biol. 2014;96(5):713-27.
[25] García E, Jones GE, Machesky LM, Antón IM. WIP: WASP-interacting proteins at invadopodia and podosomes. Eur J Cell Biol. 2012; 91(11-12):869-77.
[26] Rohatgi R, Ho HY, Kirschner MW. Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. J Cell Biol. 2000; 150(6):1299-310.
[27] Helgeson LA, Nolen BJ. Mechanism of synergistic activation of Arp2/3 complex by cortactin and N-WASP. Elife. 2013; 2:e00884.
[28] Fang XJ, Jiang H, Zhu YQ, Zhang LY, Fan QH, Tian Y. Doxorubicin induces drug resistance and expression of the novel CD44st via NF-κB in human breast cancer MCF-7 cells. Oncol Rep. 2014; 31(6):2735-42.
[29] Wan X, Hou J, Liu S, Zhang Y, Li W, Zhang Y, Ding Y. Estrogen Receptor α Mediates Doxorubicin Sensitivity in Breast Cancer Cells by Regulating E-Cadherin. Front Cell Dev Biol. 2021; 9:583572.
[30] Rasouli A, Aliebrahimi S, Montazeri V, Ghahremani MH, Ostad SN. Combination effect of doxorubicin and HIF inhibitor on MCF-7 CD44+/CD24- subpopulation cells in hypoxic condition. Braz J Pharm Sci. 2022; 58:581-10.
[31] Mizielska A, Dziechciowska I, Szczepański R, Cisek M, Dąbrowska M, Ślężak J, Kosmalska I, Rymarczyk M, Wilkowska K, Jacczak B, Totoń E, Lisiak N, Kopczyński P, Rubiś B. Doxorubicin and Cisplatin Modulate miR-21, miR-106, miR-126, miR-155 and miR-199 Levels in MCF7, MDA-MB-231 and SK-BR-3 Cells That Makes Them Potential Elements of the DNA-Damaging Drug Treatment Response Monitoring in Breast Cancer Cells-A Preliminary Study. Genes (Basel). 2023; 14(3):702.