Superior role for p53 in DNA breakage compared with parthanatos by tumor microenvironment of breast cancer cells

Subject Areas : Developmental biology of plants and animals , development and differentiation in microorganisms

Shahrzad Chitgaran ¹ , Shadi Rabiee ² , Mahmood Barati ³ , Parisa Hayat ⁴ , Abdolhamid Angaji ⁵ , Shima Tavakol ⁶

1 - Department of Cell and Molecular Biology, Faculty of Biological Science, Kharazmi University, Tehran, Iran
2 - Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran
3 - Department of Biotechnology, Faculty of Paramedicine, Iran University of Medical Sciences, Tehran, Iran
4 - Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
5 - Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, P.O. Box: 15719-14911, Tehran, Iran
6 - Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran

Received: 2024-11-13 Accepted : 2025-03-20 Published : 2025-03-10

Keywords: Breast Neoplasms, Conditioned Culture Media, Parthanatos, Apoptosis, DNA Damage, Acidosis,

Abstract :

Background: Cells are continuously exposed to various cellular stresses. Over 50% of human cancers carry mutations in the p53 gene, therefore p53 has been considered a critical tumor suppressor. Upon DNA damage, p53 accumulates in the cellular nucleus through post-translational modifications.

Method and materials: In this study, the impact of an acidic and conditioned MDA-MBA231 breast cancer environment on the pathways of apoptosis and parthanatos in normal fibroblast cells was examined. The expression of Bcl2, Bax genes involved in apoptosis, PARP1, PARG genes involved in parthanatos, and p53 protein was evaluated. Moreover, the electrophoresis method was used to study DNA breakage in normal, acidic, conditioned, and acidic plus conditioned media.

Results: The results showed that the level of DNA damage in acidic and conditioned media was less than that in conditioned media and higher than that in normal and acidic media. Although conditioned media induced less Bax and Bax/Bcl2 ratio than other media in fibroblasts, there was no significant difference between the Bax/Bcl2 ratio in other groups as the apoptosis index. PARP1 and PARG gene levels were the lowest in conditioned plus acidic media, and p53 was downregulated in conditioned media.

 

Conclusion: Parthanatos and the level of Bax were not responsible for DNA damage in conditioned media, whereas a decrease in p53 was partly associated with poor DNA repair under stress conditions in cancer cells in normal fibroblasts.

References:

[1] S. Rabiee, S. Tavakol, M. Barati, and M.T. Joghataei, Autophagic, apoptotic, and necrotic cancer cell fates triggered by acidic pH microenvironment. Journal of cellular physiology 234 (2019) 12061-12069.
[2] P.A. Jones, and S.B. Baylin, The epigenomics of cancer. Cell 128 (2007) 683-692.
[3] S. Tavakol, M.R. Nikpour, E. Hoveizi, B. Tavakol, S.M. Rezayat, M. Adabi, S. Shajari Abokheili, and M.J.J.o.n.r. Jahanshahi, Investigating the effects of particle size and chemical structure on cytotoxicity and bacteriostatic potential of nano hydroxyapatite/chitosan/silica and nano hydroxyapatite/chitosan/silver; as antibacterial bone substitutes. 16 (2014) 1-13.
[4] E. Baxter, K. Windloch, F. Gannon, and J.S. Lee, Epigenetic regulation in cancer progression. Cell & bioscience 4 (2014) 1-11.
[5] S. Rabiee, E. Hoveizi, M. Barati, A. Salehzadeh, M.T. Joghataei, and S.J.P.o. Tavakol, Cancer cells same as zombies reprogram normal cells via the secreted microenvironment. 18 (2023) e0288003.
[6] R. Baghban, L. Roshangar, R. Jahanban-Esfahlan, K. Seidi, A. Ebrahimi-Kalan, M. Jaymand, S. Kolahian, T. Javaheri, and P. Zare, Tumor microenvironment complexity and therapeutic implications at a glance. Cell Communication and Signaling 18 (2020) 1-19.
[7] D. Hanahan, and L.M. Coussens, Accessories to the crime: functions of cells recruited to the tumor microenvironment. Cancer cell 21 (2012) 309-322.
[8] J.W. Wojtkowiak, D. Verduzco, K.J. Schramm, and R.J. Gillies, Drug resistance and cellular adaptation to tumor acidic pH microenvironment. Molecular pharmaceutics 8 (2011) 2032-2038.
[9] G. Hao, Z.P. Xu, and L. Li, Manipulating extracellular tumour pH: an effective target for cancer therapy. RSC advances 8 (2018) 22182-22192.
[10] J.S. Fang, R.D. Gillies, and R.A. Gatenby, Adaptation to hypoxia and acidosis in carcinogenesis and tumor progression, Seminars in cancer biology, Elsevier, 2008, pp. 330-337.
[11] Y. Kato, T. Maeda, A. Suzuki, and Y. Baba, Cancer metabolism: new insights into classic characteristics. Japanese Dental Science Review 54 (2018) 8-21.
[12] C. Corbet, and O. Feron, Tumour acidosis: from the passenger to the driver's seat. Nature Reviews Cancer 17 (2017) 577-593.
[13] A. Som, S. Bloch, J.E. Ippolito, and S. Achilefu, Acidic extracellular pH of tumors induces octamer-binding transcription factor 4 expression in murine fibroblasts in vitro and in vivo. Scientific reports 6 (2016) 1-6.
[14] D. Yang, T. Shu, H. Zhao, Y. Sun, W. Xu, and G. Tu, Knockdown of macrophage migration inhibitory factor (MIF), a novel target to protect neurons from parthanatos induced by simulated post-spinal cord injury oxidative stress. Biochemical and Biophysical Research Communications 523 (2020) 719-725.
[15] F. Sanchez-vega, Oncogenic Signaling Pathways in The Cancer Article Oncogenic Signaling Pathways in The Cancer Genome Atlas. (2018). 173(2): 321–337.e10.
[16] L. Galluzzi, I. Vitale, S.A. Aaronson, J.M. Abrams, D. Adam, P. Agostinis, E.S. Alnemri, L. Altucci, I. Amelio, and D.W. Andrews, Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death & Differentiation 25 (2018) 486-541.
[17] S. CZENE, M. TIBÄCK, and M. HARMS-RINGDAHL, pH-dependent DNA cleavage in permeabilized human fibroblasts. Biochemical Journal 323 (1997) 337-341.
[18] K.S. Carswell, and E.T. Papoutsakis, Extracellular pH affects the proliferation of cultured human T cells and their expression of the interleukin-2 receptor. Journal of immunotherapy 23 (2000) 669-674.
[19] R. Martínez-Zaguilán, E.A. Seftor, R.E. Seftor, Y.-W. Chu, R.J. Gillies, and M.J. Hendrix, Acidic pH enhances the invasive behavior of human melanoma cells. Clinical & experimental metastasis 14 (1996) 176-186.
[20] L. Borsi, E. Balza, B. Gaggero, G. Allemanni, and L. Zardi, The Alternative Splicing Pattern of the Tenascin-C Pre-mRNA Is Controlled by the Extracellular pH (∗). Journal of Biological Chemistry 270 (1995) 6243-6245.
[21] F. Skenderi, S. Vranic, and I. Damjanov, Regulated cell death in diagnostic histopathology. International Journal of Developmental Biology 59 (2015) 149-158.
[22] R.S. Wong, Apoptosis in cancer: from pathogenesis to treatment. Journal of experimental & clinical cancer research 30 (2011) 1-14.
[23] R. Venkatadri, T. Muni, A. Iyer, J. Yakisich, and N. Azad, Role of apoptosis-related miRNAs in resveratrol-induced breast cancer cell death. Cell death & disease 7 (2016) e2104-e2104.
[24] S. Hassab El-Nabi, and Y. Elhassaneen, Detection of DNA damage, molecular apoptosis and production of home-made ladder by using simple techniques. Biotechnology 7 (2008) 514-522.
[25] S. Rabiee Motmaen, S. Tavakol, M.T. Joghataei, and M. Barati, Acidic pH derived from cancer cells as a double‐edged knife modulates wound healing through DNA repair genes and autophagy. International Wound Journal 17 (2020) 137-148.
[26] T. Akoto, and S. Saini, Role of exosomes in prostate cancer metastasis. International journal of molecular sciences 22 (2021) 3528.
[27] F. Hadi, S. Tavakkol, S. Laurent, V. Pirhajati, S.R. Mahdavi, A. Neshastehriz, and A. Shakeri‐Zadeh, Combinatorial effects of radiofrequency hyperthermia and radiotherapy in the presence of magneto‐plasmonic nanoparticles on MCF‐7 breast cancer cells. Journal of cellular physiology 234 (2019) 20028-20035.
[28] Y. Zhou, L. Liu, S. Tao, Y. Yao, Y. Wang, Q. Wei, A. Shao, and Y. Deng, Parthanatos and its associated components: promising therapeutic targets for cancer. Pharmacological Research 163 (2021) 105299.
[29] B. Arneth, Tumor microenvironment. Medicina 56 (2019) 15.
[30] H. Park, J. Lyons, T. Ohtsubo, and C. Song, Acidic environment causes apoptosis by increasing caspase activity. British journal of cancer 80 (1999) 1892-1897.
[31] S.A. Andrabi, T.M. Dawson, and V.L. Dawson, Mitochondrial and nuclear cross talk in cell death: parthanatos. Annals of the New York Academy of Sciences 1147 (2008) 233-241.
[32] F. Moosavi, B. Hassani, and O. Firuzi, Combination of PARP inhibitor, Olaparib, with DNAdamaging chemotherapeutic agents. Koomesh 25 (1402) 522.
[33] P. Gao, D. Wang, M. Liu, S. Chen, Z. Yang, J. Zhang, H. Wang, Y. Niu, W. Wang, and J. Yang, DNA methylation-mediated repression of exosomal miR-652-5p expression promotes oesophageal squamous cell carcinoma aggressiveness by targeting PARG and VEGF pathways. PLoS genetics 16 (2020) e1008592.
[34] P. Bertheau, J. Lehmann-Che, M. Varna, A. Dumay, B. Poirot, R. Porcher, E. Turpin, L.-F. Plassa, A. de Roquancourt, and E. Bourstyn, p53 in breast cancer subtypes and new insights into response to chemotherapy. The Breast 22 (2013) S27-S29.
[35] H.E. Marei, A. Althani, N. Afifi, A. Hasan, T. Caceci, G. Pozzoli, A. Morrione, A. Giordano, and C. Cenciarelli, p53 signaling in cancer progression and therapy. Cancer cell international 21 (2021) 1-15.
[36] J. Huang, Current developments of targeting the p53 signaling pathway for cancer treatment. Pharmacology & Therapeutics 220 (2021) 107720.
[37] F. Zamani, S. Oraee-Yazdani, L. Langroudi, and S.M.J.K. Hashemi, Role of mesenchymal stem cells in growth and progression of cancer and prospective potentials in cancer therapy. Koomesh 24 (2024) 1-25.