Annexin V FITC conjugated as a radiation toxicity indicator in lymphocytes following radiation overexposure in radiotherapy programs

Tavakoli, Mohammad Bagher; Kheirollahi, Majid; Kiani, Ali; Kazemi, Mohammad; Haghjooy Javanmard, Shaghayegh; Mohebat, Leili

Annexin V FITC conjugated as a radiation toxicity indicator in lymphocytes following radiation overexposure in radiotherapy programs

Authors

¹ Department of Medical Physics, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

² Department of Genetics, Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non-communicable Disease, Isfahan University of Medical Sciences, Isfahan, Iran

³ Department of Physiology, Isfahan University of Medical Sciences, Isfahan, Iran

⁴ Department of Oncology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Following human radiation exposure in hospital or accidents, dose assessments are of prime importance in radiation accidents. These issues are of continuing importance with respect to socioeconomic policy relating to the industrial and medical uses of ionizing radiation, and also for risk assessment among people who are occupationally exposed to low and/or high linear energy transfer (LET) radiation, such as astronauts, pilots, stewardesses, nuclear power plant workers, and victims of radiation accidents.
Materials and Methods: In this study, an assay for assessing radiation dose based on the induction of apoptosis in human T-lymphocytes was done to examine T-lymphocyte cells isolated from the fresh blood of 16volunteers, cultured and exposed to gamma rays. Radiation-induced apoptosis (RIA) was assessed by flow cytometric identification of cells displaying apoptosis-associated DNA condensation.
Results: Dose-response experiments showed that at 2Gy dose level of radiotherapy programs, the RIA frequency was significantly above control. Apoptotic levels significantly depend on the dose of radiation rather than the donor.
Conclusion: The results demonstrate the potential use of this assay as a biological indicator of radiation toxicity, optimizing patient dose in radiotherapy and biological dosimetry process.

Keywords

References

1.	vanEngeland M, Nieland LJ, Ramaekers FC, Schutte B, Reutelingsperger CP. Annexin V-affinity assay: A review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry 1998;31:1-9.
2.	Bordón E, Henríquez Hernández LA, Lara PC, Pinar B, Fontes F, Rodríguez Gallego C, et al. Prediction of clinical toxicity in localized cervical carcinoma by radio-induced apoptosis study in peripheral blood lymphocytes (PBLs). Radiat Oncol 2009;4:58.
3.	Ilyenko I, Lyaskivska O, Bazyka D. Analysis of relative telomere length and apoptosis in humans exposed to ionising radiation. Exp Oncol 2011;33:235-8.
4.	Hooker DJ, Mobarok M, Anderson JL, Rajasuriar R, Gray LR, Ellett AM, et al. A new way of measuring apoptosis by absolute quantitation of inter-nucleosomally fragmented genomic DNA. Nucleic Acids Res 2012;40:e113.
5.	Bordón E, Henríquez-Hernández LA, Lara PC, Ruíz A, Pinar B, Rodríguez-Gallego C, et al. Prediction of clinical toxicity in locally advanced head and neck cancer patients by radio-induced apoptosis in peripheral blood lymphocytes (PBLs). Radiat Oncol 2010;5:4.
6.	Kakizaki T, Hamada N, Sakashita T, Wada S, Hara T, Funayama T, et al. Vulnerability of feline T-lymphocytes to charged particles. J Vet Med Sci 2007;69:605-9.
7.	International Atomic Energy Agency (IAEA). Handling of radiation accidents. In: Dolphin GW, editor. Biological Dosimetry with Particular References to Chromosome Aberration Analysis: A Review of Methods. Vienna, Austria:; 1969. p. 215-24.
8.	Menz R, Andres R, Larsson B, Ozsahin M, Trott K, Crompton NE. Biological dosimetry: The potential use of radiation-induced apoptosis in human T-lymphocytes. Radiat Environ Biophys 1997;36:175-81.
9.	Hosokawa Y, Sakakura Y, Tanaka L, Okumura K, Yajima T, Kaneko M. Radiation induced apoptosis is independent of caspase-8 but dependent on cytochrome C and the caspase-9cascade in human leukemia HL60 cells. J Radiat Res 2005;46:293-303.
10.	Amaral A. Trends in biological dosimetry: An overview. Braz Arch Biol Technol 2002;45:119-24.
11.	Pinar B, Henríquez-Hernández LA, Lara PC, Bordon E, Rodriguez-Gallego C, Lloret M, et al. Radiation induced apoptosis and initial DNA damage are inversely related in locally advanced breast cancer patients. Radiat Oncol 2010;5:85.
12.	Hodge S, Hodge G, Holmes M, Reynolds PN. Increased peripheral blood T-cell apoptosis and decreased Bcl-2 in chronic obstructive pulmonary disease. Immunol Cell Biol 2005;83:160-6.
13.	Schnarr K, Boreham D, Sathya J, Julian J, Dayes IS. Radiation-induced lymphocyte apoptosis to predict radiation therapy late toxicity in prostate cancer patients. Int J Radiat Oncol Biol Phys 2009;74:1424-30.
14.	Ishihara S, Iinuma H, Fukushima Y, Akahane T, Horiuchi A, Shimada R, et al. Radiation-induced apoptosis of peripheral blood lymphocytes is correlated with histological regression of rectal cancer in response to preoperative chemoradiotherapy. Ann Surg Oncol 2012;19:1192-8.
15.	Prieto A, Díaz D, Barcenilla H, García-Suárez J, Reyes E, Monserrat J, et al. Apoptotic rate: A new indicator for the quantification of the incidence of apoptosis in cell cultures. Cytometry 2002;48:185-93.
16.	Zhang HM, Nie JS, Li X, Niu Q. Characteristic analysis of peripheral blood mononuclear cell apoptosis in coke oven workers. J Occup Health 2012;54:44-50.
17.	Zhang G, Gurtu V, Kain SR, Yan G. Early detection of apoptosis using a fluorescent conjugate of annexin V. Biotechniques 1997;23:525-31.
18.	Hingorani R, Deng J, Elia J, McIntyre C, Mittar D. Detection of Apoptosis Using the BD Annexin V FITC Assay on the BD FACS Verse System: Flow cytometry base protocol. USA, San Diego, CA: BD Biosciences; 2011. p. 1-12.

Advanced Biomedical Research