Analysis of chromosomal radiosensitivity of healthy BRCA2 mutation carriers and non-carriers in BRCA families with the G2 micronucleus assay

Baert,Annelot; Depuydt,Julie; Van Maerken,Tom; Poppe,Bruce; Malfait,Fransiska; Van Damme,Tim; De Nobele,Sylvia; Perletti,Gianpaolo; De Leeneer,Kim; Claes,Kathleen B.M.; Vral,Anne

doi:10.3892/or.2017.5407

Analysis of chromosomal radiosensitivity of healthy BRCA2 mutation carriers and non-carriers in BRCA families with the G2 micronucleus assay

Authors:
- Annelot Baert
- Julie Depuydt
- Tom Van Maerken
- Bruce Poppe
- Fransiska Malfait
- Tim Van Damme
- Sylvia De Nobele
- Gianpaolo Perletti
- Kim De Leeneer
- Kathleen B.M. Claes
- Anne Vral
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Affiliations: Department of Basic Medical Sciences, Ghent University, B-9000 Ghent, Belgium, Department of Pediatrics and Medical Genetics, Ghent University, B-9000 Ghent, Belgium, Center for Medical Genetics, Ghent University Hospital, B-9000 Ghent, Belgium
Published online on: January 25, 2017 https://doi.org/10.3892/or.2017.5407
Pages: 1379-1386
Copyright: © Baert et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Breast cancer risk drastically increases in individuals with a heterozygous germline BRCA1 or BRCA2 mutation, while it is estimated to equal the population risk for relatives without the familial mutation (non-carriers). The aim of the present study was to use a G2 phase-specific micronucleus assay to investigate whether lymphocytes of healthy BRCA2 mutation carriers are characterized by increased radiosensitivity compared to controls without a family history of breast/ovarian cancer and how this relates to healthy non-carrier relatives. BRCA2 is active in homologous recombination, a DNA damage repair pathway, specifically active in the late S/G2 phase of the cell cycle. We found a significantly increased radiosensitivity in a cohort of healthy BRCA2 mutation carriers compared to individuals without a familial history of breast cancer (P=0.046; Mann-Whitney U test). At the individual level, 50% of healthy BRCA2 mutation carriers showed a radiosensitive phenotype (radiosensitivity score of 1 or 2), whereas 83% of the controls showed no radiosensitivity (P=0.038; one-tailed Fisher's exact test). An odds ratio of 5 (95% CI, 1.07-23.47) indicated an association between the BRCA2 mutation and radiosensitivity in healthy mutation carriers. These results indicate the need for the gentle use of ionizing radiation for either diagnostic or therapeutic use in BRCA2 mutation carriers. We detected no increased radiosensitivity in the non-carrier relatives.

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1	Roy R, Chun J and Powell SN: BRCA1 and BRCA2: Different roles in a common pathway of genome protection. Nat Rev Cancer. 12:68–78. 2011. View Article : Google Scholar : PubMed/NCBI
2	Bernholtz S, Laitman Y, Kaufman B, Shimon-Paluch S and Friedman E: Phenocopy breast cancer rates in Israeli BRCA1 BRCA2 mutation carrier families: Is the risk increased in non-carriers? Breast Cancer Res Treat. 132:669–673. 2012. View Article : Google Scholar : PubMed/NCBI
3	Domchek SM, Gaudet MM, Stopfer JE, Fleischaut MH, Powers J, Kauff N, Offit K, Nathanson KL and Robson M: Breast cancer risks in individuals testing negative for a known family mutation in BRCA1 or BRCA2. Breast Cancer Res Treat. 119:409–414. 2010. View Article : Google Scholar : PubMed/NCBI
4	Harvey SL, Milne RL, McLachlan SA, Friedlander ML, Birch KE, Weideman P, Goldgar D, Hopper JL and Phillips KA: kConFab Investigators: Prospective study of breast cancer risk for mutation negative women from BRCA1 or BRCA2 mutation positive families. Breast Cancer Res Treat. 130:1057–1061. 2011. View Article : Google Scholar : PubMed/NCBI
5	Korde LA, Mueller CM, Loud JT, Struewing JP, Nichols K, Greene MH and Mai PL: No evidence of excess breast cancer risk among mutation-negative women from BRCA mutation-positive families. Breast Cancer Res Treat. 125:169–173. 2011. View Article : Google Scholar : PubMed/NCBI
6	Kurian AW, Gong GD, John EM, Johnston DA, Felberg A, West DW, Miron A, Andrulis IL, Hopper JL, Knight JA, et al: Breast cancer risk for noncarriers of family-specific BRCA1 and BRCA2 mutations: Findings from the Breast Cancer Family Registry. J Clin Oncol. 29:4505–4509. 2011. View Article : Google Scholar : PubMed/NCBI
7	Nielsen HR, Petersen J, Krogh L, Nilbert M and Skytte AB: No evidence of increased breast cancer risk for proven noncarriers from BRCA1 and BRCA2 families. Fam Cancer. 15:523–528. 2016. View Article : Google Scholar : PubMed/NCBI
8	Smith A, Moran A, Boyd MC, Bulman M, Shenton A, Smith L, Iddenden R, Woodward ER, Lalloo F, Maher ER, et al: Phenocopies in BRCA1 and BRCA2 families: Evidence for modifier genes and implications for screening. J Med Genet. 44:10–15. 2007. View Article : Google Scholar : PubMed/NCBI
9	Vos JR, De Bock GH, Teixeira N, van der Kolk DM, Jansen L, Mourits MJE and Oosterwijk JC: Proven non-carriers in BRCA families have an earlier age of onset of breast cancer. Eur J Cancer. 49:2101–2106. 2013. View Article : Google Scholar : PubMed/NCBI
10	Evans DGR, Ingham SL, Buchan I, Woodward ER, Byers H, Howell A, Maher ER, Newman WG and Lalloo F: Increased rate of phenocopies in all age groups in BRCA1/BRCA2 mutation kindred, but increased prospective breast cancer risk is confined to BRCA2 mutation carriers. Cancer Epidemiol Biomarkers Prev. 22:2269–2276. 2013. View Article : Google Scholar : PubMed/NCBI
11	Pijpe A, Andrieu N, Easton DF, Kesminiene A, Cardis E, Noguès C, Gauthier-Villars M, Lasset C, Fricker JP, Peock S, et al: GENEPSO; EMBRACE; HEBON: Exposure to diagnostic radiation and risk of breast cancer among carriers of BRCA1/2 mutations: Retrospective cohort study (GENE-RAD-RISK). BMJ. 345:e56602012. View Article : Google Scholar : PubMed/NCBI
12	Baert A, Depuydt J, Van Maerken T, Poppe B, Malfait F, Storm K, van den Ende J, Van Damme T, De Nobele S, Perletti G, et al: Increased chromosomal radiosensitivity in asymptomatic carriers of a heterozygous BRCA1 mutation. Breast Cancer Res. 18:522016. View Article : Google Scholar : PubMed/NCBI
13	Andrieu N, Easton DF, Chang-Claude J, Rookus MA, Brohet R, Cardis E, Antoniou AC, Wagner T, Simard J, Evans G, et al: Effect of chest X-rays on the risk of breast cancer among BRCA1/2 mutation carriers in the international BRCA1/2 carrier cohort study: A report from the EMBRACE, GENEPSO, GEO-HEBON, and IBCCS Collaborators' Group. J Clin Oncol. 24:3361–3366. 2006. View Article : Google Scholar : PubMed/NCBI
14	Lecarpentier J, Noguès C, Mouret-Fourme E, Stoppa-Lyonnet D, Lasset C, Caron O, Fricker JP, Gladieff L, Faivre L, Sobol H, et al: GENEPSO: Variation in breast cancer risk with mutation position, smoking, alcohol, and chest X-ray history, in the French National BRCA1/2 carrier cohort (GENEPSO). Breast Cancer Res Treat. 130:927–938. 2011. View Article : Google Scholar : PubMed/NCBI
15	Narod SA, Lubinski J, Ghadirian P, Lynch HT, Moller P, Foulkes WD, Rosen B, Kim-Sing C, Isaacs C, Domchek S, et al: Hereditary Breast Cancer Clinical Study Group: Screening mammography and risk of breast cancer in BRCA1 and BRCA2 mutation carriers: A case-control study. Lancet Oncol. 7:402–406. 2006. View Article : Google Scholar : PubMed/NCBI
16	Giannakeas V, Lubinski J, Gronwald J, Moller P, Armel S, Lynch HT, Foulkes WD, Kim-Sing C, Singer C, Neuhausen SL, et al: Mammography screening and the risk of breast cancer in BRCA1 and BRCA2 mutation carriers: A prospective study. Breast Cancer Res Treat. 147:113–118. 2014. View Article : Google Scholar : PubMed/NCBI
17	Goldfrank D, Chuai S, Bernstein JL, Ramon Y, Cajal T, Lee JB, Alonso MC, Diez O, Baiget M, Kauff ND, Offit K, et al: Effect of mammography on breast cancer risk in women with mutations in BRCA1 or BRCA2. Cancer Epidemiol Biomarkers Prev. 15:2311–2313. 2006. View Article : Google Scholar : PubMed/NCBI
18	John EM, McGuire V, Thomas D, Haile R, Ozcelik H, Milne RL, Felberg A, West DW, Miron A, Knight JA, et al: Kathleen Cuningham Foundation Consortium for Research into Familial Breast Cancer (kConFab); Whittemore AS: Diagnostic chest X-rays and breast cancer risk before age 50 years for BRCA1 and BRCA2 mutation carriers. Cancer Epidemiol Biomarkers Prev. 22:1547–1556. 2013. View Article : Google Scholar : PubMed/NCBI
19	Bernstein JL, Thomas DC, Shore RE, Robson M, Boice JD Jr, Stovall M, Andersson M, Bernstein L, Malone KE, Reiner AS, et al: WECARE Study Collaborative Group: Contralateral breast cancer after radiotherapy among BRCA1 and BRCA2 mutation carriers: A WECARE study report. Eur J Cancer. 49:2979–2985. 2013. View Article : Google Scholar : PubMed/NCBI
20	Baeyens A, Thierens H, Claes K, Poppe B, de Ridder L and Vral A: Chromosomal radiosensitivity in BRCA1 and BRCA2 mutation carriers. Int J Radiat Biol. 80:745–756. 2004. View Article : Google Scholar : PubMed/NCBI
21	Gutiérrez-Enríquez S, Ramón Y, Cajal T, Alonso C, Corral A, Carrasco P, Cornet M, Sanz J, Ribas M, Baiget M and Diez O: Ionizing radiation or mitomycin-induced micronuclei in lymphocytes of BRCA1 or BRCA2 mutation carriers. Breast Cancer Res Treat. 127:611–622. 2011. View Article : Google Scholar : PubMed/NCBI
22	Trenz K, Rothfuss A, Schütz P and Speit G: Mutagen sensitivity of peripheral blood from women carrying a BRCA1 or BRCA2 mutation. Mutat Res. 500:89–96. 2002. View Article : Google Scholar : PubMed/NCBI
23	Beroukas E, Pandis N, Giannoukakos K, Rizou E, Beroukas K, Giatromanolaki A and Koukourakis M: Increased chromosomal radiosensitivity in women carrying BRCA1/BRCA2 mutations assessed with the G2 assay. Int J Radiat Oncol Biol Phys. 76:1199–1205. 2010. View Article : Google Scholar : PubMed/NCBI
24	Becker AA, Graeser MK, Landwehr C, Hilger T, Baus W, Wappenschmidt B, Meindl A, Weber RG and Schmutzler RK: A 24-color metaphase-based radiation assay discriminates heterozygous BRCA2 mutation carriers from controls by chromosomal radiosensitivity. Breast Cancer Res Treat. 135:167–175. 2012. View Article : Google Scholar : PubMed/NCBI
25	Bolognesi C, Bruzzi P, Gismondi V, Volpi S, Viassolo V, Pedemonte S and Varesco L: Clinical application of micronucleus test: A case-control study on the prediction of breast cancer risk/susceptibility. PLoS One. 9:e1123542014. View Article : Google Scholar : PubMed/NCBI
26	Cardinale F, Bruzzi P and Bolognesi C: Role of micronucleus test in predicting breast cancer susceptibility: A systematic review and meta-analysis. Br J Cancer. 106:780–790. 2012. View Article : Google Scholar : PubMed/NCBI
27	Claes K, Depuydt J, Taylor AMR, Last JI, Baert A, Schietecatte P, Vandersickel V, Poppe B, De Leeneer K, D'Hooghe M, et al: Variant ataxia telangiectasia: Clinical and molecular findings and evaluation of radiosensitive phenotypes in a patient and relatives. Neuromolecular Med. 15:447–457. 2013. View Article : Google Scholar : PubMed/NCBI
28	Menzel T, Nähse-Kumpf V, Kousholt AN, Klein DK, Lund-Andersen C, Lees M, Johansen JV, Syljuåsen RG and Sørensen CS: A genetic screen identifies BRCA2 and PALB2 as key regulators of G2 checkpoint maintenance. EMBO Rep. 12:705–712. 2011. View Article : Google Scholar : PubMed/NCBI
29	Keimling M, Volcic M, Csernok A, Wieland B, Dörk T and Wiesmüller L: Functional characterization connects individual patient mutations in ataxia telangiectasia mutated (ATM) with dysfunction of specific DNA double-strand break-repair signaling pathways. FASEB J. 25:3849–3860. 2011. View Article : Google Scholar : PubMed/NCBI
30	Arnold K, Kim MK, Frerk K, Edler L, Savelyeva L, Schmezer P and Wiedemeyer R: Lower level of BRCA2 protein in heterozygous mutation carriers is correlated with an increase in DNA double strand breaks and an impaired DSB repair. Cancer Lett. 243:90–100. 2006. View Article : Google Scholar : PubMed/NCBI
31	Perrin-Vidoz L, Sinilnikova OM, Stoppa-Lyonnet D, Lenoir GM and Mazoyer S: The nonsense-mediated mRNA decay pathway triggers degradation of most BRCA1 mRNAs bearing premature termination codons. Hum Mol Genet. 11:2805–2814. 2002. View Article : Google Scholar : PubMed/NCBI
32	Anczuków O, Ware MD, Buisson M, Zetoune AB, Stoppa-Lyonnet D, Sinilnikova OM and Mazoyer S: Does the nonsense-mediated mRNA decay mechanism prevent the synthesis of truncated BRCA1, CHK2, and p53 proteins? Hum Mutat. 29:65–73. 2008. View Article : Google Scholar : PubMed/NCBI
33	Berger AH, Knudson AG and Pandolfi PP: A continuum model for tumour suppression. Nature. 476:163–169. 2011. View Article : Google Scholar : PubMed/NCBI
34	Guidugli L, Pankratz VS, Singh N, Thompson J, Erding CA, Engel C, Schmutzler R, Domchek S, Nathanson K, Radice P, et al: A classification model for BRCA2 DNA binding domain missense variants based on homology-directed repair activity. Cancer Res. 73:265–275. 2013. View Article : Google Scholar : PubMed/NCBI
35	Willems P, Claes K, Baeyens A, Vandersickel V, Werbrouck J, De Ruyck K, Poppe B, van den Broecke R, Makar A, Marras E, et al: Polymorphisms in nonhomologous end-joining genes associated with breast cancer risk and chromosomal radiosensitivity. Genes Chromosomes Cancer. 47:137–148. 2008. View Article : Google Scholar : PubMed/NCBI
36	Guo Z, Shu Y, Zhou H, Zhang W and Wang H: Radiogenomics helps to achieve personalized therapy by evaluating patient responses to radiation treatment. Carcinogenesis. 36:307–317. 2015. View Article : Google Scholar : PubMed/NCBI
37	Popanda O, Marquardt JU, Chang-Claude J and Schmezer P: Genetic variation in normal tissue toxicity induced by ionizing radiation. Mutat Res. 667:58–69. 2009. View Article : Google Scholar : PubMed/NCBI
38	Antoniou AC, Kuchenbaecker KB, Soucy P, Beesley J, Chen X, McGuffog L, Lee A, Barrowdale D, Healey S, Sinilnikova OM and et al; CIMBA: SWE-BRCA; HEBON; EMBRACE; GEMO Collaborators Study; kConFab Investigators: Common variants at 12p11, 12q24, 9p21, 9q31.2 and in ZNF365 are associated with breast cancer risk for BRCA1 and/or BRCA2 mutation carriers. Breast Cancer Res. 14:R332012. View Article : Google Scholar : PubMed/NCBI
39	Claes K, Poppe B, Machackova E, Coene I, Foretova L, DePaepe A and Messiaen L: Differentiating pathogenic mutations from polymorphic alterations in the splice sites of BRCA1 and BRCA2. Genes Chromosomes Cancer. 37:314–320. 2003. View Article : Google Scholar : PubMed/NCBI