DNA repair and apoptosis: Roles in radiotherapy-related acute reactions in breast cancer patients

Bahadir Batar; Tuba Mutlu; Merve Bostanci; Mustafa Akin; Matem Tuncdemir; Nuran Bese; Mehmet Guven

doi:10.14715/cmb/2018.64.4.11

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Vol. 64 No. 4: Issue 4

Issue Published : April 2, 2018

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DNA repair and apoptosis: Roles in radiotherapy-related acute reactions in breast cancer patients

https://doi.org/10.14715/cmb/2018.64.4.11

Bahadir Batar

Department of Medical Biology, Namik Kemal University Medical Faculty, Tekirdag, Turkey

Tuba Mutlu

Department of Medical Biology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey

Merve Bostanci

Department of Medical Biology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey

Mustafa Akin

Department of Clinical Radiation Oncology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey

Matem Tuncdemir

Department of Medical Biology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey

Nuran Bese

Department of Clinical Radiation Oncology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey

Mehmet Guven

Department of Medical Biology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey

Corresponding Author(s) : Bahadir Batar

bahadirbatar@gmail.com

Cellular and Molecular Biology, Vol. 64 No. 4: Issue 4
Article Published : March 31, 2018

Abstract

Normal tissue reactions are therapy limiting factor for the effectiveness of the radiotherapy in cancer patients. DNA repair and apoptosis are estimated to be critical players of adverse effects in response to radiotherapy. Our aim was to define the association of DNA repair (ERCC1 and XPC) and apoptotic (BCL2, CASP3 and NFKB1) gene expression, DNA damage levels, apoptosis changes and DNA repair gene variations with the risk of acute side effects in breast cancer patients. The study included 100 women with newly diagnosed breast cancer; an experimental case group (n=50) with acute side effects and the control group (n=50) without side effects. Gene expression was analyzed by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). Micronucleus (MN) and 8-hydroxy-2"²-deoxyguanosine (8-OHdG) assays were performed to compare the DNA damage levels. Apoptosis was examined by TDT-mediated dUTP-biotin nick end-labeling (TUNEL) staining. ERCC1 rs3212986 and XPC rs3731055 polymorphisms were genotyped by real-time PCR technique. No significantly correlation of DNA repair and apoptosis gene expression and DNA damage levels with acute side effects in response to radiotherapy. Also, there was no association between apoptosis levels and acute effects. ERCC1 rs3212986 CC genotype showed a protective effect against radiotherapy-induced acute reactions (p<0.001; OR: 0.21; 95% CI= 0.08-0.52). Our results suggest that apoptosis and DNA damage levels are not associated with acute radiosensitivity. DNA repair may affect the risk of acute reactions. Further studies are needed to validate the current findings.

Keywords

Acute side effects Apoptosis Breast cancer DNA damage DNA repair ERCC1 Radiotherapy.

Batar, B., Mutlu, T., Bostanci, M., Akin, M., Tuncdemir, M., Bese, N., & Guven, M. (2018). DNA repair and apoptosis: Roles in radiotherapy-related acute reactions in breast cancer patients. Cellular and Molecular Biology, 64(4), 64–70. https://doi.org/10.14715/cmb/2018.64.4.11

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References

Kouloulias V, Gogalis I, Zygogianni A, Armpilia C, Antypas C, Kokakis J, et al. A unique hypofractionated radiotherapy schedule with 51.3 Gy in 18 fractions three times per week for early breast cancer: outcomes including local control, acute and late skin toxicity. Breast Cancer 2017; 24:263-70.
Henriquez-Hernandez LA, Bordon E, Pinar B, Lloret M, Rodriguez-Gallego C, Lara PC. Prediction of normal tissue toxicity as part of the individualized treatment with radiotherapy in oncology patients. Surg Oncol 2012; 21:201-6.
Chang-Claude J, Popanda O, Tan XL, Kropp S, Helmbold I, von Fournier D, et al. Association between Polymorphisms in the DNA Repair Genes, XRCC1, APE1, and XPD and Acute Side Effects of Radiotherapy in Breast Cancer Patients. Clin Cancer Res 2005; 11:4802-9.
Cramers P, Filon AR, Pines A, Kleinjans JC, Mullenders LH, van Zeeland AA. Enhanced nucleotide excision repair in human fibroblasts pre-exposed to ionizing radiation. Photocem Photobiol 2012; 88:147-53.
Park CH, Bessho T, Matsunaga T, Sancar A. Purification and characterization of the XPF-ERCC1 complex of human DNA repair excision nuclease. J Biol Chem 1995; 270:22657-60.
Thoma BS, Vasquez KM. Critical DNA damage recognition functions of XPC-hHR23B and XPA-RPA in nucleotide excision repair. Mol Carcinog 2003; 38:1-13.
Wiebalk K, Schmezer P, Kropp S, Chang-Claude J, Celebi O, Debus J, et al. In vitro radiation-induced expression of XPC mRNA as a possible biomarker for developing adverse reactions during radiotherapy. Int J Cancer 2007; 121:2340-5.
Rieger KE, Hong WJ, Tusher VG, Tang J, Tibshirani R, Chu G. Toxicity from radiation therapy associated with abnormal transcriptional responses to DNA damage. Proc Natl Acad Sci USA 2004; 101:6635-40.
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; 98:1845-51.
Chua ML, 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; 53:355-64.
Sudprasert W, Navasumrit P, Ruchirawat M. Effects of low-dose gamma radiation on DNA damage, chromosomal aberration and expression of repair genes in human blood cells. Int J Hyg Environ Health 2006; 209:503-11.
Zijno A, Saini F, Crebelli R. Suitability of cryopreserved isolated lymphocytes for the analysis of micronuclei with the cytokinesis-block method. Mutagenesis 2007; 22:311-5.
Fenech M. The Cytokinesis-Block Micronucleus Technique and Its Application to Genotoxicity Studies in Human Populations. Environ Health Perspect 1993; 101:101-7.
Catena C, Parasacchi P, Conti D, Sgura A, Trenta G, Righi E, et al. Peripheral blood lymphocyte decrease and micronucleus yields during radiotherapy. Int J Radiat Biol 1997; 72:575–85.
Marková E, Somsedíková A, Vasilyev S, Pobijaková M, Lacková A, Lukaćko P, et al. DNA repair foci and late apoptosis/necrosis in peripheral blood lymphocytes of breast cancer patients undergoing radiotherapy. Int J Radiat Biol 2015; 91:934-45.
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.
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.
Ozsahin M, Crompton NE, Gourgou S, Kramar A, Li L, Shi Y, et al. CD4 and CD8 T-lymphocyte apoptosis can predict radiation-induced late toxicity: A prospective study in 399 patients. Clin Cancer Res 2005; 11:7426-33.
Finnon P, Kabacik S, MacKay A, Raffy C, A'Hern R, Owen R, et al. Correlation of in vitro lymphocyte radiosensitivity and gene expression with late normal tissue reactions following curative radiotherapy for breast cancer. Radiother Oncol 2012; 105:329-36.
Beaton LA, Ferrarotto C, Marro L, Samiee S, Malone S, Grimes S, et al. Chromosome Damage and Cell Proliferation Rates in In Vitro Irradiated Whole Blood as Markers of Late Radiation Toxicity After Radiation Therapy to the Prostate. Int J Radiat Oncol Biol Phys 2013; 85:1346-52.
Barber JB, Burrill W, Spreadborough AR, Levine E, Warren C, Kiltie AE, et al. Relationship between in vitro chromosomal radiosensitivity of peripheral blood lymphocytes and the expression of normal tissue damage following radiotherapy for breast cancer. Radiother Oncol 2000; 55:179-86.
Chua ML, Somaiah N, A'Hern R, Davies S, Gothard L, Yarnold J, et al. Residual DNA and chromosomal damage in ex vivo irradiated blood lymphocytes correlated with late normal tissue response to breast radiotherapy. Radiother Oncol 2011; 99:362-6.
Padjas A, Kedzierawski P, Florek A, Kukolowicz P, Kuszewski T, GóÅºdz S, et al. Comparative analysis of three functional predictive assays in lymphocytes of patients with breast and gynaecological cancer treated by radiotherapy. J Contemp Brachytherapy 2012; 4:219-26.
Batar B, Guven G, Eroz S, Bese NS, Guven M. Decreased DNA repair gene XRCC1 expression is associated with radiotherapy-induced acute side effects in breast cancer patients. Gene 2016; 582:33-7.
Evans MD, Saparbaev M, Cooke MS. DNA repair and the origins of urinary oxidized 2'- deoxyribonucleosides. Mutagenesis 2010; 25:433-42.
Roszkowski K, Olinski R. Urinary 8-oxoguanine as a predictor of survival in patients undergoing radiotherapy. Cancer Epidemiol Biomarkers Prev 2012; 2012:629-35.
Haghdoost S, Svoboda P, Ingemar N. Can 8-oxo-dg be used as a predictor for individual radiosensitivity? Int J Radiat Oncol Biol Phys 2001; 50:405-10.
Werbrouck J, De Ruyck K, Beels L, Vral A, Van Eijkeren M, De Neve W, et al. Prediction of late normal tissue complications in RT treated gynaecological cancer patients: Potential of the gamma-H2AX foci assay and association with chromosomal radiosensitivity. Oncol Rep 2010; 23:571-8.
Vandevoorde C, Depuydt J, Veldeman L, De Neve W, Sebastií N, Wieme G, et al. In vitro cellular radiosensitivity in relationship to late normal tissue reactions in breast cancer patients: a multi-endpoint case-control study. Int J Radiat Biol 2016; 17:1-14.
Lavin MF, Shiloh Y. The genetic defect in ataxia-telangiectasia. Annu Rev Immunol 1997; 15:177202.
Carney JP. Chromosomal breakage syndromes. Curr Opin Immunol 1999; 11:443-7.
Turesson I, Nyman J, Holmberg E, Odén A. Prognostic factors for acute and late skin reactions in radiotherapy patients. Int J Radiat Oncol Biol Phys 1996; 36:1065-75.
Hümmerich J, Werle-Schneider G, Popanda O, Celebi O, Chang-Claude J, et al. Constitutive mRNA expression of DNA repair-related genes as a biomarker for clinical radio-resistance: A pilot study in prostate cancer patients receiving radiotherapy. Int J Radiat Biol 2006; 82:593-604.
Svensson JP, Stalpers LJ, Esveldt-van Lange RE, Franken NA, Haveman J, Klein B, et al. Analysis of gene expression using gene sets discriminates cancer patients with and without late radiation toxicity. PLoS Med 2006; 3:e422.
Mangoni M, Bisanzi S, Carozzi F, Sani C, Biti G, Livi L,et al. Association between genetic polymorphisms in the XRCC1, XRCC3, XPD, GSTM1, GSTT1, MSH2, MLH1, MSH3, and MGMT genes and radiosensitivity in breast cancer patients. Int J Radiat Oncol Biol Phys 2011; 81:52-8.
Vral A, Willems P, Claes K, Poppe B, Perletti ABG, Thierens H. Combined effect of polymorphisms in Rad51 and XRCC3 on breast cancer risk and chromosomal radiosensitivity. Mol Med Rep 2011; 185:901-12.
Sánchez-Suárez P, Ostrosky-Wegman P, Gallegos-Hernández F, Peñarroja-Flores R, Toledo-García J, Bravo JL, et al. DNA damage in peripheral blood lymphocytes in patients during combined chemotherapy for breast cancer. Mutat Res 2008; 640:8-15.
Andreassen CN, Alsner J, Overgaard M, Overgaard J. Prediction of normal tissue radiosensitivity from polymorphisms in candidate genes. Radiother Oncol 2003; 69:127-35.
Sterpone S, Cornetta T, Padua L, Mastellone V, Giammarino D, Testa A, et al. DNA repair capacity and acute radiotherapy adverse effects in Italian breast cancer patients. Mutat Res 201; 684:43-8.
Sakano S, Hinoda Y, Sasaki M, Wada T, Matsumoto H, Eguchi S, et al. Nucleotide excision repair gene polymorphisms may predict acute toxicity in patients treated with chemoradiotherapy for bladder cancer. Pharmacogenomics. 2010; 11:1377-87.

References

Kouloulias V, Gogalis I, Zygogianni A, Armpilia C, Antypas C, Kokakis J, et al. A unique hypofractionated radiotherapy schedule with 51.3 Gy in 18 fractions three times per week for early breast cancer: outcomes including local control, acute and late skin toxicity. Breast Cancer 2017; 24:263-70.

Henriquez-Hernandez LA, Bordon E, Pinar B, Lloret M, Rodriguez-Gallego C, Lara PC. Prediction of normal tissue toxicity as part of the individualized treatment with radiotherapy in oncology patients. Surg Oncol 2012; 21:201-6.

Chang-Claude J, Popanda O, Tan XL, Kropp S, Helmbold I, von Fournier D, et al. Association between Polymorphisms in the DNA Repair Genes, XRCC1, APE1, and XPD and Acute Side Effects of Radiotherapy in Breast Cancer Patients. Clin Cancer Res 2005; 11:4802-9.

Cramers P, Filon AR, Pines A, Kleinjans JC, Mullenders LH, van Zeeland AA. Enhanced nucleotide excision repair in human fibroblasts pre-exposed to ionizing radiation. Photocem Photobiol 2012; 88:147-53.

Park CH, Bessho T, Matsunaga T, Sancar A. Purification and characterization of the XPF-ERCC1 complex of human DNA repair excision nuclease. J Biol Chem 1995; 270:22657-60.

Thoma BS, Vasquez KM. Critical DNA damage recognition functions of XPC-hHR23B and XPA-RPA in nucleotide excision repair. Mol Carcinog 2003; 38:1-13.

Wiebalk K, Schmezer P, Kropp S, Chang-Claude J, Celebi O, Debus J, et al. In vitro radiation-induced expression of XPC mRNA as a possible biomarker for developing adverse reactions during radiotherapy. Int J Cancer 2007; 121:2340-5.

Rieger KE, Hong WJ, Tusher VG, Tang J, Tibshirani R, Chu G. Toxicity from radiation therapy associated with abnormal transcriptional responses to DNA damage. Proc Natl Acad Sci USA 2004; 101:6635-40.

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; 98:1845-51.

Chua ML, 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; 53:355-64.

Sudprasert W, Navasumrit P, Ruchirawat M. Effects of low-dose gamma radiation on DNA damage, chromosomal aberration and expression of repair genes in human blood cells. Int J Hyg Environ Health 2006; 209:503-11.

Zijno A, Saini F, Crebelli R. Suitability of cryopreserved isolated lymphocytes for the analysis of micronuclei with the cytokinesis-block method. Mutagenesis 2007; 22:311-5.

Fenech M. The Cytokinesis-Block Micronucleus Technique and Its Application to Genotoxicity Studies in Human Populations. Environ Health Perspect 1993; 101:101-7.

Catena C, Parasacchi P, Conti D, Sgura A, Trenta G, Righi E, et al. Peripheral blood lymphocyte decrease and micronucleus yields during radiotherapy. Int J Radiat Biol 1997; 72:575–85.

Marková E, Somsedíková A, Vasilyev S, Pobijaková M, Lacková A, Lukaćko P, et al. DNA repair foci and late apoptosis/necrosis in peripheral blood lymphocytes of breast cancer patients undergoing radiotherapy. Int J Radiat Biol 2015; 91:934-45.

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.

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.

Ozsahin M, Crompton NE, Gourgou S, Kramar A, Li L, Shi Y, et al. CD4 and CD8 T-lymphocyte apoptosis can predict radiation-induced late toxicity: A prospective study in 399 patients. Clin Cancer Res 2005; 11:7426-33.

Finnon P, Kabacik S, MacKay A, Raffy C, A'Hern R, Owen R, et al. Correlation of in vitro lymphocyte radiosensitivity and gene expression with late normal tissue reactions following curative radiotherapy for breast cancer. Radiother Oncol 2012; 105:329-36.

Beaton LA, Ferrarotto C, Marro L, Samiee S, Malone S, Grimes S, et al. Chromosome Damage and Cell Proliferation Rates in In Vitro Irradiated Whole Blood as Markers of Late Radiation Toxicity After Radiation Therapy to the Prostate. Int J Radiat Oncol Biol Phys 2013; 85:1346-52.

Barber JB, Burrill W, Spreadborough AR, Levine E, Warren C, Kiltie AE, et al. Relationship between in vitro chromosomal radiosensitivity of peripheral blood lymphocytes and the expression of normal tissue damage following radiotherapy for breast cancer. Radiother Oncol 2000; 55:179-86.

Chua ML, Somaiah N, A'Hern R, Davies S, Gothard L, Yarnold J, et al. Residual DNA and chromosomal damage in ex vivo irradiated blood lymphocytes correlated with late normal tissue response to breast radiotherapy. Radiother Oncol 2011; 99:362-6.

Padjas A, Kedzierawski P, Florek A, Kukolowicz P, Kuszewski T, GóÅºdz S, et al. Comparative analysis of three functional predictive assays in lymphocytes of patients with breast and gynaecological cancer treated by radiotherapy. J Contemp Brachytherapy 2012; 4:219-26.

Batar B, Guven G, Eroz S, Bese NS, Guven M. Decreased DNA repair gene XRCC1 expression is associated with radiotherapy-induced acute side effects in breast cancer patients. Gene 2016; 582:33-7.

Evans MD, Saparbaev M, Cooke MS. DNA repair and the origins of urinary oxidized 2'- deoxyribonucleosides. Mutagenesis 2010; 25:433-42.

Roszkowski K, Olinski R. Urinary 8-oxoguanine as a predictor of survival in patients undergoing radiotherapy. Cancer Epidemiol Biomarkers Prev 2012; 2012:629-35.

Haghdoost S, Svoboda P, Ingemar N. Can 8-oxo-dg be used as a predictor for individual radiosensitivity? Int J Radiat Oncol Biol Phys 2001; 50:405-10.

Werbrouck J, De Ruyck K, Beels L, Vral A, Van Eijkeren M, De Neve W, et al. Prediction of late normal tissue complications in RT treated gynaecological cancer patients: Potential of the gamma-H2AX foci assay and association with chromosomal radiosensitivity. Oncol Rep 2010; 23:571-8.

Vandevoorde C, Depuydt J, Veldeman L, De Neve W, Sebastií N, Wieme G, et al. In vitro cellular radiosensitivity in relationship to late normal tissue reactions in breast cancer patients: a multi-endpoint case-control study. Int J Radiat Biol 2016; 17:1-14.

Lavin MF, Shiloh Y. The genetic defect in ataxia-telangiectasia. Annu Rev Immunol 1997; 15:177202.

Carney JP. Chromosomal breakage syndromes. Curr Opin Immunol 1999; 11:443-7.

Turesson I, Nyman J, Holmberg E, Odén A. Prognostic factors for acute and late skin reactions in radiotherapy patients. Int J Radiat Oncol Biol Phys 1996; 36:1065-75.

Hümmerich J, Werle-Schneider G, Popanda O, Celebi O, Chang-Claude J, et al. Constitutive mRNA expression of DNA repair-related genes as a biomarker for clinical radio-resistance: A pilot study in prostate cancer patients receiving radiotherapy. Int J Radiat Biol 2006; 82:593-604.

Svensson JP, Stalpers LJ, Esveldt-van Lange RE, Franken NA, Haveman J, Klein B, et al. Analysis of gene expression using gene sets discriminates cancer patients with and without late radiation toxicity. PLoS Med 2006; 3:e422.

Mangoni M, Bisanzi S, Carozzi F, Sani C, Biti G, Livi L,et al. Association between genetic polymorphisms in the XRCC1, XRCC3, XPD, GSTM1, GSTT1, MSH2, MLH1, MSH3, and MGMT genes and radiosensitivity in breast cancer patients. Int J Radiat Oncol Biol Phys 2011; 81:52-8.

Vral A, Willems P, Claes K, Poppe B, Perletti ABG, Thierens H. Combined effect of polymorphisms in Rad51 and XRCC3 on breast cancer risk and chromosomal radiosensitivity. Mol Med Rep 2011; 185:901-12.

Sánchez-Suárez P, Ostrosky-Wegman P, Gallegos-Hernández F, Peñarroja-Flores R, Toledo-García J, Bravo JL, et al. DNA damage in peripheral blood lymphocytes in patients during combined chemotherapy for breast cancer. Mutat Res 2008; 640:8-15.

Andreassen CN, Alsner J, Overgaard M, Overgaard J. Prediction of normal tissue radiosensitivity from polymorphisms in candidate genes. Radiother Oncol 2003; 69:127-35.

Sterpone S, Cornetta T, Padua L, Mastellone V, Giammarino D, Testa A, et al. DNA repair capacity and acute radiotherapy adverse effects in Italian breast cancer patients. Mutat Res 201; 684:43-8.

Sakano S, Hinoda Y, Sasaki M, Wada T, Matsumoto H, Eguchi S, et al. Nucleotide excision repair gene polymorphisms may predict acute toxicity in patients treated with chemoradiotherapy for bladder cancer. Pharmacogenomics. 2010; 11:1377-87.

Author biographies is not available.