Volume 8, Issue 2, December 2020, Page: 76-81
Determination of Endogenous Radioresistance Biomarkers on HeLa Cells
Mamadou Soumboundou, Laboratory of Biophysics, Department of Biology and Functional Explorations, Thies University, Thies, Senegal; Diagnostic Laboratory, Children Hospital of Diamniadio, Dakar, Senegal; Laboratory of Pharmaceutical Physics, Pharmacy Department, University Cheikh Anta Diop of Dakar (UCAD), Dakar, Senegal
Marie Thérèse Aloy, Molecular and Cellular Radiobiology Laboratory, University of Claude Bernard, Villeurbanne, France
Macoura Gadji, Service of Hematology, National Centre of Blood Transfusion (CNTS), Department of Applied Biological and Pharmaceutical Sciences, Faculty of Medicine, Pharmacy and Odontology (FMPO), University Cheikh Anta Diop of Dakar (UCAD), Dakar, Senegal; Research Institute in Oncology & Haematology (RIOH), Cancer Care Manitoba (CCMB), Department of Physiology & Pathophysiology, University of Manitoba (UFM), Winnipeg, Manitoba, Canada
Celine Malesys, Molecular and Cellular Radiobiology Laboratory, University of Claude Bernard, Villeurbanne, France
Gora Mbaye, Laboratory of Pharmaceutical Physics, Pharmacy Department, University Cheikh Anta Diop of Dakar (UCAD), Dakar, Senegal
Ahmadou Dem, Cancer Institute, Department of surgery, University Cheikh Anta Diop of Dakar (UCAD), Dakar, Senegal
Mounibe Diarra, Laboratory of Pharmaceutical Physics, Pharmacy Department, University Cheikh Anta Diop of Dakar (UCAD), Dakar, Senegal
Claire Rodriguez-Lafrasse, Molecular and Cellular Radiobiology Laboratory, University of Claude Bernard, Villeurbanne, France
Received: Oct. 18, 2019;       Accepted: Nov. 27, 2019;       Published: Dec. 28, 2020
DOI: 10.11648/j.ejb.20200802.18      View  40      Downloads  14
Abstract
The Genes such as carbonic anhydrase IX (CAIX), hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) have been suggested as hypoxic biomarkers in cancer. Indeed, these endogenous biomarkers have been shown to have stronger prognostic value response after treatment by irradiation. However, inconsistent results suggest that factors other than oxygen influence their expression. This present study deciphers the level of expression of different radioresistance biomarkers in both normoxia and hypoxia conditions followed by irradiation of human ovarian tumor cell lines (uterine cervix squamous cell carcinoma (HeLa). HeLa cells were submitted to hypoxia (1% O2) conditions in a Thermo Scientific Heracell i CO2 incubator. The cells were subjected to two doses 4-10 Gy irradiation and re-incubate in their starting conditions for 4 hours, then fixed in 4% paraformaldehyde for 20 min. Protein expressions were assessed by immunocytochemistry staining and fluorescent images were captured by a Axio Imager Z1 fluorescence microscope with oil immersion lens at 63× magnification. In normoxia conditions there was no modification of the level of expression of the CAIX after irradiation. However, an increasing expression level of VEGF was noted. The level of expression of HIF-1 in normoxia was low compared to the other two proteins (CAIX and VEGF). Hypoxia conditions at 2% resulted in a low expression of CAIX and VEGF before and after irradiation at 10 Gy in HeLa cells. HIF-1 had a maximum expression level compared to CAIX and VEGF at 2% oxygen after irradiation in HeLa cells. As tumor hypoxia occurs in a deprived microenvironment, other environmental factors such as irradiation might interact with the effect of low oxygen concentration on gene expression. This study shows that irradiation of HeLa cells has a profound influence on the oxygen dependent induction of certain endogenous hypoxic markers as HIF-1, CAIX, and VEGF.
Keywords
Radioresistance, Biomarkers, Hypoxia, Normoxia, DNA Repair HELA Cells
To cite this article
Mamadou Soumboundou, Marie Thérèse Aloy, Macoura Gadji, Celine Malesys, Gora Mbaye, Ahmadou Dem, Mounibe Diarra, Claire Rodriguez-Lafrasse, Determination of Endogenous Radioresistance Biomarkers on HeLa Cells, European Journal of Biophysics. Vol. 8, No. 2, 2020, pp. 76-81. doi: 10.11648/j.ejb.20200802.18
Copyright
Copyright © 2020 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Bray F, Ferlay J, Soerjomataram I, Siegel R L and Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68, 6, 2018, pp 394–424.
[2]
William S; Monica A. B; Amishi B; Linus T. C and Gaffney D K. Cervical Cancer: A Global Health Crisis. Cancer, 123, 13, 2017, pp 2404-2412.
[3]
World Health Organization. Human papillomavirus (HPV) and cervical cancer. http://www.who.int/mediacentre/factsheets/fs380/en/. Accessed September 05, 2019.
[4]
Mukherjee G, Freeman A, Moore R, Kumaraswamy and Stanley MA. Biologic factors and response to radiotherapy in carcinoma of the cervix. Int J Gynecol Cancer. 11, 3, 2001, pp 187–193.
[5]
Schwarz JK, Siegel BA, Dehdashti F and Grigsby PW. Metabolic response on post-therapy FDG-PET predicts patterns of failure after radiotherapy for cervical cancer. Int J Radiat Oncol Biol Phys. 83, 2012, pp 185–190.
[6]
Pawlik TM and Keyomarsi K. Role of cell cycle in mediating sensitivity to radiotherapy. Int J Radiat Oncol Biol Phys. 59, 4, 2004, pp 928-942.
[7]
Barbara M, Pilar de la P, Feda A and Abdel K A. The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia (Auckl). 3, 2015, pp 83-92.
[8]
Ribatti D and Crivellato E. “Sprouting angiogenesis”, a reappraisal. Dev Biol., 372, 2, 2012, pp 157-165.
[9]
Bhaskar A and Tiwary B. N. Hypoxia inducible factor-1 alpha and multiple myeloma. Int. J. Adv. Res. 4, 1, 2016, pp 706-715.
[10]
Klopp AH and Eifel PJ. Biological predictors of cervical cancer response to radiation therapy. Semin Radiat Oncol. 22, 2, 2012, pp 143–150.
[11]
Christine Ellingsen, Lise Mari K. Andersen, Kanthi Galappathi and Einar K. Rofstad. Hypoxia biomarkers in squamous cell carcinoma of the uterine cervix. BMC Cancer. 15, 2015: 805.
[12]
Birner P, Schindl M, Obermair A, Plank C and Oberhuber G. Overexpression of hypoxia-inducible factor 1α is a marker for an unfavorable prognosis in early-stage invasive cervical cancer. Cancer Res. 60, 17, 2000, pp 4693–4596.
[13]
Kaluz S, Kaluzová M, Liao SY, Lerman M and Stanbridge EJ. Transcriptional control of the tumor- and hypoxia-marker carbonic anhydrase 9: a one transcription factor (HIF-1) show? Biochim Biophys Acta. 1795, 2, 2009, pp 162-172.
[14]
Loncaster JA, Harris AL, Davidson SE, Logue JP and Wycoff CC. Carbonic anhydrase (CA IX) expression, a potential new intrinsic marker of hypoxia: correlations with tumor oxygen measurements and prognosis in locally advanced carcinoma of the cervix. Cancer Res. 61, 17, 2001, pp 6394–6399.
[15]
Forker L. J, Choudhury A and Kiltie A. E. Biomarkers of Tumour Radiosensitivity and Predicting Benefit from Radiotherapy. Clinical Oncology. 27, 10, 2015, pp 561-569.
[16]
Franken NAP, Rodermond HM, Stap J, Haveman J and Van Bree C. Clonogenic assay of cells in vitro. Nat Protocols. 1, 5, 2006, pp 2315-2319.
[17]
Hanot M, Boivin A, Malesys C, Beuve M and Rodriguez-Lafrasse C. Glutathione depletion and carbon ion radiation potentiates clustered DNA lesions, cell death and prevents chromosomal changes in cancer cells progeny. Plos One 2012; 7, 11, 2012, e44367.
[18]
Dellas K, Bache M, Pigorsch SU, Taubert H and Haensgen G. Prognostic impact of HIF-1α expression in patients with definitive radiotherapy for cervical cancer. Strahlenther Onkol. 184, 3, 2008, pp 169-174.
[19]
Hasmim M, Messai Y, Noman M. Z and Chouaib S. L’hypoxie tumorale: un déterminant clé de la réactivité stromale et de la réponse antitumorale. Med Sci (Paris). 2014, 30 (4), p 422-428.
[20]
Jinsil S. Challenge and hope in radiotherapy of hepatocellular carcinoma. Yonsei Med J. 50, 5, 2009, pp 601–612.
[21]
Moeller BJ and Dewhirst MW. HIF-1 and tumour radiosensitivity. Br J Cancer. 95, 1, 2006, pp 1–5.
[22]
Bertout JA, Patel SA and Simon MC. The impact of O2 availability on human cancer. Nat Rev Cancer. 8, 12, 2008, pp 967–975.
[23]
Moeller BJ, Dreher MR, Rabbani ZN, Schroeder T and Dewhirst MW. Pleiotropic effects of HIF-1 blockade on tumor radiosensitivity. Cancer Cell. 8, 2, 2005, pp 99-110.
[24]
Burri P, Djonov V, Aebersold DM, Lindel K and Gruber G. Significant correlation of hypoxia-inducible factor-1α with treatment outcome in cervical cancer treated with radical radiotherapy. Int J Radiat Oncol Biol Phys. 56, 2, 2003, pp 494–501.
[25]
Birner P, Schindl M, Obermair A, Breitenecker G and Oberhuber G. Expression of hypoxia-inducible factor 1α in epithelial ovarian tumors: its impact on prognosis and on response to chemotherapy. Clin Cancer Res. 7, 6, 2001, pp 1661–1668.
[26]
Daniele G, Alfredo B, Maria P. B, Leticia C and Stephen B. Fox. Hypoxia-Inducible Factor-1α Expression Predicts a Poor Response to Primary Chemoendocrine Therapy and Disease-Free Survival in Primary Human Breast Cancer. Clin Cancer Res. 12, 15, 2006, pp 4562-4568.
[27]
Hutchison GJ, Valentine HR, Loncaster JA, Davidson SE and West CM. Hypoxia-inducible factor 1alpha expression as an intrinsic marker of hypoxia: correlation with tumor oxygen, pimonidazole measurements, and outcome in locally advanced carcinoma of the cervix. Clin Cancer Res. 10, 24, 2004, pp 8405–8412.
[28]
Yuefen Z, Chao C, Motoko O-M, Yun K and Tomio I. The suppression of hypoxia inducible factor and vascular endothelial growth factor by siRNA does not affect the radiation sensitivity of multicellular tumor spheroids. J Radiat Res, 51, 2010, pp 47-55.
[29]
Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 3, 10, 2003, pp 721–732.
[30]
Lee S, Shin HJ, Han IO, Hong EK and Kim JY. Tumor carbonic anhydrase 9 expression is associated with the presence of lymph node metastases in uterine cervical cancer. Cancer Sci, 98, 3, 2007, p 329-333.
[31]
Pastorekova S, Zatovicova M and Pastorek J. Cancer-associated carbonic anhydrases and their inhibition. Curr Pharm Des. 14, 7, 2008, pp 685-698.
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