Bax/Bcl-2 expression ratio in prediction of response to breast cancer radiotherapy

Document Type : Original Article

Authors

1 Department of Medical Physics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Reza Radiotherapy and Oncology Center, Mashhad, Iran

3 Medical Physics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

4 Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Radiotherapy is one of the most effective modalities of cancer therapy, but clinical responses of individual patients varies considerably. To enhance treatment efficiency it is essential to implement an individual-based treatment. The aim of present study was to identify the mechanism of intrinsic apoptosis pathway on radiosensitivity and normal tissue complications caused by the radiotherapy.
Materials and Methods: Peripheral blood mononuclear cells from ten breast cancer patients were exposed to 6MV X-rays to deliver 1 and 2 Gy. Expression levels of Bax, Bcl-2, and Bax/Bcl-2 ratio were examined by relative quantitative RT-PCR. All the patients received similar tangential irradiation of the whole breast and conventional fractionation. Skin dosimetry was done by GAFChromic EBT-3 film and clinical radiosensitivity was determined using the acute reactions to radiotherapy of the skin according to Radiation Therapy Oncology Group score. All statistical analyses were performed using GraphPad Prism, version 7.01.
Results: In the in-vitro experiment, Bax and Bax/Bcl-2 ratios were significantly increased with 1 and 2 Gy doses (PP<0.0001, respectively). Herein, the notable result was a significant correlation between dose-response curve slope (as an in-vitro radiosensitivity index) and acute skin toxicity score following irradiation (as a clinical radiosensitivity index). There was no significant relationship between skin dose and reactions (P>0.05 for all patients).
Conclusion: Significant correlation between Bax/Bcl-2 ratio determined before radiation therapy and clinical response in the patients, can be used as a biomarker to identify radiosensitive individuals. However, further studies are required to validate radiation-induced apoptotic biomarkers.

Keywords

Main Subjects

References

1. Matsumura Y, Ananthaswamy HN. Toxic effects of ultraviolet radiation on the skin. Toxicol Appl Pharmacol 2004; 195:298-308.
2. Akmansu M, Erel A. Atypical acute reaction associated with radiotherapy: a case report. Radiat Med 1998; 16:379-382.
3. Holler U, Schubert T, Budach V, Trefzer U, Beyer M. Blisters - an unusual effect during radiotherapy. Strahlenther Onkol 2013; 189:977-979.
4. Stanley SE, Rao AD, Gable DL, McGrath-Morrow S, Armanios M. Radiation sensitivity and radiation necrosis in the short telomere syndromes. Int J Radiat Oncol Biol Phys 2015; 93:1115-1117.
5. Farhood B, Mahdavi SR, Emranpour MH, Mohammadi Asl K, Nekoui N, Knaup C. Skin reaction in radiation therapy for breast cancer. Iran J Med Phys 2014; 11:316-321.
6. Obexer P, Ausserlechner MJ. X-linked inhibitor of apoptosis protein–a critical death resistance regulator and therapeutic target for personalized cancer therapy. Front Oncol 2014; 4:197.
7. Brengues M, Lapierre A, Bourgier C, Pèlegrin A, Özsahin M, Azria D. T lymphocytes to predict radiation-induced late effects in normal tissues. Expert Rev Mol Diagn 2017; 17:119-127.
8. Gatti RA, Painter RB. Ataxia-telangiectasia: Springer Science & Business Media; 2013.
9. Birkeland AC, Auerbach AD, Sanborn E, Parashar B, Kuhel WI, Chandrasekharappa SC, et al. Postoperative clinical radiosensitivity in patients with fanconi anemia and head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg 2011; 137:930-934.
10. Murray JE, Bicknell LS, Yigit G, Duker AL, Kogelenberg M, Haghayegh S, et al. Extreme growth failure is a common presentation of ligase IV deficiency. Hum Mutat 2014; 35:76-85.
11. Bogdanova N, Feshchenko S, Schürmann P, Waltes R, Wieland B, Hillemanns P, et al. Nijmegen breakage syndrome mutations and risk of breast cancer. Int J Cancer 2008; 122:802-806.
12. Bowman KJ, Al‐Moneef MM, Sherwood BT, Colquhoun AJ, Goddard JC, Griffiths T, et al. Comet assay measures of DNA damage are predictive of bladder cancer cell treatment sensitivity in vitro and outcome in vivo. Int J Cancer 2014; 134:1102-1111.
13. Borgmann K, Hoeller U, Nowack S, Bernhard M, Röper B, Brackrock S, et al. Individual radiosensitivity measured with lymphocytes may predict the risk of acute reaction after radiotherapy. Int J Radiat Oncol Biol Phys 2008; 71:256-264.
14. Bourton EC, Plowman PN, Smith D, Arlett CF, Parris CN. Prolonged expression of the γ‐H2AX DNA repair biomarker correlates with excess acute and chronic toxicity from radiotherapy treatment. Int J Cancer 2011; 129:2928-2934.
15. Chua MLK, Horn S, Somaiah N, Davies S, Gothard L, A’Hern R, et al. DNA double-strand break repair and induction of apoptosis in ex vivo irradiated blood lymphocytes in relation to late normal tissue reactions following breast radiotherapy. Radiat Environ Biophys 2014; 53:355-364.
16. Forrester HB, Li J, Leong T, McKay MJ, Sprung CN. Identification of a radiation sensitivity gene expression profile in primary fibroblasts derived from patients who developed radiotherapy-induced fibrosis. Radiother Oncol 2014; 111:186-193.
17. Mumbrekar KD, Fernandes DJ, Goutham HV, Sharan K, Vadhiraja BM, Satyamoorthy K, et al. Influence of double-strand break repair on radiation therapy-induced acute skin reactions in breast cancer patients. Int J Radiat Oncol Biol Phys 2014; 88:671-676.
18. Oeck S, Al-Refae K, Riffkin H, Wiel G, Handrick R, Klein D, et al. Activating Akt1 mutations alter DNA double strand break repair and radiosensitivity. Sci Rep 2017; 7:42700.
19. Brown JM, Wilson G. Apoptosis genes and resistance to cancer therapy: what does the experimental and clinical data tell us? Cancer Biol Ther 2003; 2:477-490.
20. Azimian H, Bahreyni-Toossi MT, Rezaei AR, Rafatpanah H, Hamzehloei T, Fardid R. Up-regulation of Bcl-2 expression in cultured human lymphocytes after exposure to low doses of gamma radiation. J Med Phys 2015; 40:38-44.
21. Reed JC, Pellecchia M. Apoptosis-based therapies for hematologic malignancies. Blood 2005; 106:408-418.
22. Reed J, Jurgensmeier J, Matsuyama S. Bcl-2 family proteins and mitochondria. Biochim Biophys Acta 1998; 1366:127-137.
23. Scarlett JL, Murphy MP. Release of apoptogenic proteins from the mitochondrial intermembrane space during the mitochondrial permeability transition. FEBS letters 1997; 418:282-286.
24. Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, et al. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 1997; 275:1129-1132.
25. Liu J, Huang R, Lin D, Peng J, Wu X, Lin Q, et al. Expression of survivin and bax/bcl-2 in peroxisome proliferator activated receptor-γ ligands induces apoptosis on human myeloid leukemia cells in vitro. Ann Oncol 2005; 16:455-459.
26. Farnebo M, Bykov VJ, Wiman KG. The p53 tumor suppressor: a master regulator of diverse cellular processes and therapeutic target in cancer. Biochem Biophys Res Commun 2010; 396:85-89.
27. Chipuk JE, Kuwana T, Bouchier-Hayes L, Droin NM, Newmeyer DD, Schuler M, et al. Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 2004; 303:1010-1014.
28. Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin H, Liebermann DA, et al. Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene 1994; 9:1799-1805.
29. Cox JD, Stetz J, Pajak TF. Toxicity criteria of the radiation therapy oncology group (RTOG) and the European organization for research and treatment of cancer (EORTC). Int J Radiat Oncol Biol Phys 1995; 31:1341-1346.
30. Mackey TJ, Borkowski A, Amin P, Jacobs SC, Kyprianou N. bcl-2/bax ratio as a predictive marker for therapeutic response to radiotherapy in patients with prostate cancer. Urology 1998; 52:1085-1090.
31. Huber R, Braselmann H, Geinitz H, Jaehnert I, Baumgartner A, Thamm R, et al. Chromosomal radiosensitivity and acute radiation side effects after radiotherapy in tumour patients-a follow-up study. Radiat Oncol 2011; 6:32- 39.
32. Chang HJ, Jung KH, Kim DY, Jeong S-Y, Choi HS, Kim YH, et al. Bax, a predictive marker for therapeutic response to preoperative chemoradiotherapy in patients with rectal carcinoma. Hum Pathol 2005; 36:364-371.
33. Garcia-Barros M, Paris F, Cordon-Cardo C, Lyden D, Rafii S, Haimovitz-Friedman A, et al. Tumor response to radiotherapy regulated by endothelial cell apoptosis. Science 2003; 300:1155-1159.
34. Qin C, Chen X, Bai Q, Davis MR, Fang Y. Factors associated with radiosensitivity of cervical cancer. Anticancer Res 2014; 34:4649-4656.
35. Daidone M, Luisi A, Veneroni S, Benini E, Silvestrini R. Clinical studies of bcl-2 and treatment benefit in breast cancer patients. Endocrine-related cancer 1999; 6:61-68.
36. Yuan W, Xiaoyun H, Haifeng Q, Jing L, Weixu H, Ruofan D, et al. MicroRNA-218 enhances the radiosensitivity of human cervical cancer via promoting radiation induced apoptosis. Int J Med Sci 2014; 11:691-696.
37. Harima Y, Harima K, Shikata N, Oka A, Ohnishi T, Tanaka Y. Bax and Bcl-2 expressions predict response to radiotherapy in human cervical cancer. J Cancer Res Clin Oncol 1998; 124:503-510.
38. Lee J-U, Hosotani R, Wada M, Doi R, Kosiba T, Fujimoto K, et al. Role of Bcl-2 family proteins (Bax, Bcl-2 and Bcl-X) on cellular susceptibility to radiation in pancreatic cancer cells. Eur J Cancer 1999; 35:1374-1380.
39. Matsumoto H, Wada T, Fukunaga K, Yoshihiro S, Matsuyama H, Naito K. Bax to Bcl-2 ratio and Ki-67 index are useful predictors of neoadjuvant chemoradiation therapy in bladder cancer. Jpn J Clin Oncol 2004; 34:124-130.
40. Hernández LAH, Lara PC, Pinar B, Bordón E, Gallego CR, Bilbao C, et al. Constitutive gene expression profile segregates toxicity in locally advanced breast cancer patients treated with high-dose hyperfractionated radical radiotherapy. Radiat Oncol 2009; 4:17.
41. Mayer C, Popanda O, Greve B, Fritz E, Illig T, Eckardt-Schupp F, et al. A radiation-induced gene expression signature as a tool to predict acute radiotherapy-induced adverse side effects. Cancer Lett 2011; 302:20-28.
42. Torres-Roca JF, Eschrich S, Zhao H, Bloom G, Sung J, McCarthy S, et al. Prediction of radiation sensitivity using a gene expression classifier. Cancer Res 2005; 65:7169-7176.
 
43. Lu X-X, Wu H, Sun X-M, Wang Y-L, Huang R, Xu J. Correlation between regulation of Cox-2 gene expression and radiosensitivity mechanism of esophageal cancer. Int J Clin Exp Pathol 2016; 9:2083-2090.
44. Mohammadi M, Islamian JP, Karami H, Oladghaffari M, Farajollahi A, Nejati-Koshki K. Role of HDM2 gene in radio-sensitivity of esophageal cancer cell lines to irradiation. Int J Cancer Manag 2017;10:1- 6.
45. Badie C, Dziwura S, Raffy C, Tsigani T, Alsbeih G, Moody J,           et al. Aberrant CDKN1A transcriptional response associates with abnormal sensitivity to radiation treatment. Br J Cancer 2008; 98:1845-1851.
46. Wei H, Wen-Ming C, Jun-Bo J. Plasma miR-145 as a                       novel biomarker for the diagnosis and radiosensitivity prediction of human cervical cancer. J Int Med Res 2017; 45:1054- 1060.
47. Sarosiek KA, Letai A. Directly targeting the mitochondrial pathway of apoptosis for cancer therapy using BH3 mimetics–recent successes, current challenges and future promise. FEBS J 2016; 283:3523-3533.
48. Kunogi H, Sakanishi T, Sueyoshi N, Sasai K. Prediction of radiosensitivity using phosphorylation of histone H2AX and apoptosis in human tumor cell lines. Int J Radiat Biol 2014; 90:587-593.
Iranian Journal of Basic Medical Sciences
Volume 21, Issue 3
March 2018
Pages 325-332

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