Molecular Detection of BRCA1 and BRCA2 Mutation in Breast Cancer Patients in Erbil Using PCR Techniques

Gihan H.S. Hamad1, Jaladet M. S. Jubrael2 & Salah A. Ali3

1 Dept. of Biology, College of Education, University of Mosul, Iraq

2 Scientific Research Center, College of Science, Duhok University, Kurdistan Region in Iraq

3 Histopathology Department, Hawler Medical University,Kurdistan Region in Iraq.

DOI: https://doi.org/10.17656/jzs.10533

Abstract

Breast Cancer is one of the leading causes of death in women and It is known that one quarter of women are expected to develop breast cancer at some ages of their lives. Mutations of breast cancer susceptibility genes 1 and 2 (BRCA1 and BRCA2) are the most well recognized gene mutations responsible for an increased risk of developing breast cancer. This investigation was carried out on 50 patients (all were females) who were diagnosed as breast cancer patients attended Nanakali and Rzgari Hospital in Erbil. This study was planned to detect the probable occurrence of three founder breast cancer mutations in female patients by the use of PCR technique. The outcome of genetic analysis indicated that the percentage of 185del AG mutation was 50 (1 patients) whereas, the percentage of 5382ins C mutation was 50 (4 patients) in BRCA1 gene and the third mutation 6174del T in BRCA2 not detected in any patient from 50 samples. The study demonstrated that the frequency of BRCA1 mutation (10%) was higher than BRCA2 (0%) in this sample of Kurdistan women with breast cancer.

Key Words: BRCA1, BRCA2, breast cancer, Erbil, mutation, PCR technique.

References

[1] Abeliovich, D., Kaduri, L., Lerer, I., et al. "The founder mutations 185delAG and 5382insC in BRCA1  and 6174delT in BRCA2 ap-pear in 60% of ovarian cancer and 30% of early-onset breast cancer patients  among Ashkenazi women", Am J. Hum Genet, Vol. 60, pp. 505-514. (1997).

[2] Ah Mew, N., Hamel, N., Galvez, M., Al-Saffar, M., Foulkes, "W. D. Haplotype analysis of a BRCA1:0   185delAG mutation in a Chilean family supports its Ashkenazi origins",Clin Genet, Vol. 62, pp. 151-156. (2002).

[3] Andersen, T. I., Borresen, A. L., Moller, P. A. "A common BRCA1 mutation in Norwegian breast and ovarian cancer families?", Amer. J. Hum. Genet. Vol. 59,  No. 2, pp. 486-487. (1996).

[4] Anglian Breast Cancer Study Group. "Prevalence and penetrance of BRCA1 and BRCA2 mutations in a population-based series of breast cancer cases", Anglian Breast Cancer Study Group. Br J. Cancer, Vol. 83, pp. 1301–1308. (2000).

[5] Antoniou, A., Pharoah, P. D., Narod, S., et al."Average risks of breast and ovarian cancer associated with BRCA1 or BRCA2 mutations detected in case series unselected for family history: a combined analysis of 22 studies", Am J. Hum Genet, Vol. 72, pp. 1117–1130. (2003).

[6] Arason, A., Jonasdottir, A., Barkardottir, R. B., et al."A population study of muta-tions and LOH at breast cancer gene loci in tumours from sister pairs: two recurrent mutations seem to account for all BRCA1/BRCA2 linked breast cancer in Iceland", J. Med Genet,Vol. 35, pp. 446-449. (1998).

[7] Atour, M. F. and Al-Kayed, S. A. "Mutation analysis of the breast cancer gene BRCA1 among breast cancer Jordanian females", Saudi Med J., Vol. 25, No. 1, pp. 60-63. (2004).

[8] Bar-Sade, R. B., Kruglikova, A., Modan, B., et al."The 185delAG BRCA1 mutation originated before the dispersion of Jews in the Diaspora and is not limited to Ashkenazim", Hum Mol Genet, Vol. 7, pp. 801- 805. (1998).

[9] Brose, M. S., Rebbeck, T. R., Calzone, K. A., et al. "Cancer risk estimates for   BRCA1 mutation carriers identified in a risk evaluation program", J. Natl Cancer Inst., Vol. 94, pp. 1365-1372.( 2002).

[10]Burcos, T., Cimponeriu, D. , Ion, D. A., et al. "Analysis of several BRCA1 and BRCA2 mutations in a Hospital-based Series of unselected breast Cancer Cases", Chirurgia, Vol. 108, No. 4, pp. 468-472, (2013).

[11]Chan, P. C., Wong, B. Y., Ozcelik, H., Cole, D. E. "Simple and rapid de-tection of BRCA1 and BRCA2 mutations by multiplex mutage-netically separated PCR",ClinChem, Vol. 45.pp. 1285-1287. (1999).

[12]Chen, J. J., Silver, D., Cantor, S., Livingston, D. M., Scully, R. "BRCA1, BRCA2 and RAD51 operate in a common DNA damage response pathway",Cancer Res, Vol. 59, pp. 1752– 1756. (1999).

[13]Chen, P. L., Chen, C. F., Chen, Y., Xiao, J., Sharp, Z. D., Lee, W. H. "The BRC repeats in BRCA2 are critical for RAD51 binding and resistance to methyl methane sulfonate treatment", Proc Nat AcadSci USA, Vol. 95, pp. 5287–5292. (1998).

[14]Collins, N., McManus, R., Wooster, R., et al. "Consistent loss of the wild type allele in breast cancers from a family linked to the BRCA2 gene on chromosome 13q12-13", Oncogene, Vol. 10, pp. 1673-1675, (1995).

[15]Cox, A., Dunning, A. M., Garcia-Closas, M., et al. "A common coding variant in CASP8 is associated with breast cancer risk", Nat Genet, Vol. 39, pp. 352–358. (2007).

[16]Couch, F. J., Deshano, M. L., Blackwood, M. A., et al."BRCA1mutations in women attending clinics that evaluate the risk of breast cancer", N. Engl J. Med, Vol. 336, pp. 1409 –1415. (1997).

[17]Diez, O., Domenech, M., Alonso, M. C., et al. "Identification of the 185delAG BRCA1 mutation in a Spanish Gypsy population", Hum Genet, Vol. 103, pp. 707-708. (1998).

[18]Diez, O., Osorio, A., Robledo, M., et al. "Prevalence of BRCA1 and BRCA2 Jewish mutations in  Spanish breast cancer patients", Br J Cancer, Vol. 79, pp. 1302-1303. (1999).

[19]Easton, D., McGuffog, L., Thompson, D., Dunning, A., Tee, L., et al."CHEK2*1100delC and susceptibility to breast cancer: A collaborative analysis involving 10,860 breast cancer cases and 9,065 controls from 10 studies", Am J Hum Genet,Vol. 74, pp. 1175–1182. (2004).

[20]El Gezeery, A., Mahmud, N., Moustafa, A., Mahrous, H., Mahmoud, H., Abd El-Menam, N. "BRCA1 gene mutation in familial breast cancer", Turkish J. Cancer, Vol. 38, pp. 167–174. (2008).

[21]FitzGerald, M. G., MacDonald, D. J., Krainer, M., et al. "Germ-line BRCA1 mutations in Jewish and non-Jewish women with early-onset breast cancer",N Engl J. Med.,Vol. 334, pp. 143 – 149. (1996).

[22]Ford, D., Easton, D. F., Stratton, M., et al."Genetic heterogeneity and penetrance analysis of the  BRCA1and BRCA2 genes in breast cancer families", The Breast Cancer Linkage Consortium. Am J. Hum Genet, Vol. 62, pp. 676–689. (1998).

[23]Friedman, L. S., Szabo, C. I., Ostermeyer, E. A., et al. "Novel inherited mutations and variable expressivity of BRCA1 alleles, including the founder mutation 185delAG in Ashkenazi Jewish families", Am J Hum Genet, Vol. 57, pp. 284-297. (1995).

[24]Gowen, L. C., Johnson, B. L., Latour, A. M., Sulik, K. K., Koller, B. H. "Brca1ciency results in early        embryonic lethality characterized byneuroepithelial abnormalities", Nat Genet, Vol. 12, No. 2, pp. 191-194. (1996).

[25]Grzybowska, E., Siemińska, M., Zientek, H., et al. "Germline mutations in the BRCA1 gene predisposing to breast and ovarian cancers in Upper Silesia population", ActaBiochim. Pol., Vol. 49, pp. 351–356. (2002).

[26]Guran, S., Ozet, A., Dede, M., Gille, J. J. and Yenen, M. C. "Hereditary breast cancer syndromes in a Turkish population. Results of molecular germline analysis", Cancer Genet.Cytogenet, Vol. 160, pp.164–168. (2005).

[27]Hakansson, S., Johannsson, O., Johannsson, U., et al. "Moderate frequency of BRCA1 and BRCA2 germ-line mutations in Scandinavian familial breast cancer", Amer. J. Hum. Genet., Vol. 60, No. 5, pp. 1068-1078. (1996).

[28]Hall, J. M., Lee, M. K., Newman, B., et al. "Linkage of early-onset familial breast cancer to chromosome 17q21", Science, Vol. 250, pp. 1684–1689. (1990).

[29]Hartge, P., Struewing, J. P., Wacholder, S., Brody, L. C., Tucker, M. A. "The prevalence of common BRCA1 and BRCA2 mutations among Ashkenazi Jews", Am. J.Hum Genet, Vol.64, pp.963-970, (1999).

[30]Huusko, P., Paakkonen, K., Launonen, V., et al. "Evidence of founder mutations in Finnish breast cancer families", Am J. Hum Genet, Vol. 62, pp. 1544-1548. (1998).

[31]Ikeda, N., Miyoshi, Y., Yoneda, K., et al. "Frequency of BRCA1 and BRCA2 germline mutations in Japanese breast cancer families", Int. J. Cancer, Vol. 91, pp. 83–98. (2001).

[32]Janiszewska, H., Haus, O., Lauda-Swieciak, A., et al. "Frequency of three BRCA1 gene founder mutations in breast/ovarian cancer families from the Pomerania-Kujawy region of Poland",Clin Genet,Vol. 64, No. 6, pp. 502-508. (2003).

[33]Jara, L., Ampuero, S., Seccia, L., et al. "Frecuencia de la mutación 185delAG en el gen BRCA1 en mujereschilenas sanas con antecedentsfamiliares de cáncer de mama",Revmed Chile, Vol. 130, pp.1113-1123. (2002).

[34]Johannsson, O., Ostermeyer, E. A., Hakansson, S., et al. "Founding BRCA1 mutations in hereditary breast and ovarian cancer in southern Sweden", Amer.J. Hum.Genet, Vol.58, No.3, pp 441–450, (1996). 

[35]Koifman, S., Jorge Koifman, R. "Breast cancer mortality among Ashkenazi Jewish women in São Paulo and Porto Alegre", Brazil. Breast Cancer Res., Vol. 3, No. 4, pp. 270-275. (2001).

[36]Levine, A. J. "The tumor suppressor genes",Annu Rev Biochem, Vol. 62, pp. 623-651. (1993).

[37]Levy-Lahad, E., Catane, R., Eisenberg, S., et al. "Founder BRCA1 and BRCA2 mutations in Ashkenazi Jews in Israel: frequency and differential pen-etrance in ovarian cancer and in breast-ovarian cancer  fam-ilies",Am J. Hum Genet, Vol. 60, pp. 1059 – 1067. (1997).

[38]Llort, G., Peris, M. and Blanco, I. "Hereditary breast and ovarian cancer: primary and secondary prevention for BRCA1 and BRCA2 mutation carriers", Med. Clin., Vol. 128, pp. 468-476. (2007).

[39]Lynch, H.T., Silva, E. Snyder, C., Lynch, J. F. "Hereditary breast cancer:   Part   I. diagnosing hereditary  breast  cancer syndromes",The B. J., Vol. 14, pp. 3–13. (2008).

[40]Mathew, A., Pandey, M., Rajan, B. "Do younger woman with non-metastatic and non-inflammatory breast carcinoma have poor prognosis?", World J SurgOncol, Vol. 2, No. 2., pp. 1-7. (2004).

[41]Mehdipour, P., Hosseini-Asl, S., Savabi-E, A., Habibi, L., Alvandi, E., Atri, M. "Low Frequency of  185delAG Founder Mutation of BRCA1 Gene in Iranian Breast Cancer Patients",  J Cancer Molecules,Vol. 2, pp. 123 – 127. (2006).

[42]Meuth, M. "The structure of mutation in mammalian cells", BiochimBiophysActa,Vol. 1032, pp. 1-17. (1990).

[43]Miki, Y., Swensen, J., Shattuck-Eidens, D., et al. "A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1", Science, Vol.266, pp. 66-71. (1994).

[44]Miki, Y., Swensen, J., Shattuck Eiden, D. and Futreal, P.A. "A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1", Science, Vol.266, pp. 66-71. (1997).

[45]Mullineaux, L. G., Castellano, T. M., Shaw, J., et al. "Identification of germline 185delAG BRCA1 mutations in non-Jewish Americans of Spanish ancestry from the San Luis Valley, Colorado", Cancer,Vol.98, pp. 597–602. (2003).

[46]Neuhausen, S., Gilewski, T., Norton, L., et al. "Recurrent BRCA2 6174delT mutations in Ashkenazi Jewish women affected by breast cancer", Nature Genet, Vol. 13, pp. 126–128. (1996).

[47]Neuhausen, S. L. "Founder populations and their uses for breast cancer genetics", Breast Cancer Res., Vol.2, No. 2, pp. 77-81. (2000).

[48]Neuhausen, S. L. and Marshall, C. J. "Loss of heterozygosity in familial tumors from three BRCA1-linked kindreds", Cancer Res, Vol. 54, pp. 6069-6072. (1994).

[49]Osorio, A., Robledo, M., Albertos, J., et al. "Molecular analysis of the six most recurrent mutations in the BRCA1 gene in 87 Spanish breast/ovarian cancer families", Cancer Lett. Vol.123, pp 153-158, (1998).

[50]Parkin, D. M., Bray, F., Ferlay, J., Pisani, P. "Global Cancer Statistics 2002", Cancer J. Clin., Vol. 55, No. 2, pp. 74–108. (2005).

[51]Parvin, G. D., Sankaran, S. "The BRCA1 E3 ubiquitin ligase controls centrosome dynamics", Cell Cycle,Vol.5, pp. 1946–1950. (2006).

[52]Peelen, T., van Vliet, M., Petrij-Bosch, A., et al. "A high proportion of novel mutations in BRCA1 with strong founder effects among Dutch and Belgian hereditary breast and ovarian cancer families",  Am J. Hum Genet, Vol. 60, pp. 1041-1049. (1997).

[53]Perrin-Vidoz, L., Sinilnikova, O. M., Stoppa-Lyonnet, D., Lenoir, G. M., Mazoyer, S. "The nonsense-mediated mRNA decay pathway triggers degradation of most BRCA1 mRNAs bearing premature termination codons", Hum Mol Genet., Vol. 11, No. 23, pp. 2805-2814. (2002).

[54]Phelan, C. M., Borg, A., Cuny, M., et al. "Consortium study on 1280 breast carcinomas: allelic loss on chromosome 17 targets subregions associated with family history and clinical parameters", Cancer Res, Vol. 58, pp. 1004 -1012. (1998).

[55]Pohlreich, P., Zikan, M., Stribrna, J., Kleibl, Z., Janatova, M., Kotlas, J. "High proportion of recurrent germline mutations in the BRCA1 gene in breast and ovarian cancer patients from the Prague area", Breast Cancer Res, Vol.7, No. 5, pp. 728-736. (2005).

[56]Polyak, K. "Breast cancer genes discovery". Web site   www.expertreviews.org.(2002).

[57]Rahman, N., Seal, S., Thompson, D., et al. "PALB2, which encodes a BRCA2-interacting protein, is a breast cancer susceptibility gene", Nat Genet, Vol. 39, pp. 165–167. (2007).

[58]Ramus, S. J., Kote-Jarai, Z., Friedman, L. S., et al. "Analysis of BRCA1 and BRCA2 mutations in Hungarian families with breast or breast-ovarian cancer", Am J Hum Genet., Vol.60, No. 5, pp. 1242-1246. (1997).

[59]Rashid, M. U., Zaidi, A., Torres, D., et al. "Prevalence of BRCA1 and BRCA2 mutations in Pakistani breast and ovarian cancer patients", Int. J. Cancer, Vol. 119, pp. 2832–2839. (2006).

[60]Renwick, A., Thompson, D., Seal, S., et al. "ATM mutations that cause ataxia-telangiectasia are breast cancer susceptibility alleles", Nat Genet, Vol. 38, pp. 873–875. (2006).

[61]Rio, P. G., Pernin, D., Bay, J. O., et al. "Loss of heterozygosity of BRCA1, BRCA2 and ATM genes in sporadic invasive ductal breast carcinoma",Int J Oncol, Vol. 13, pp. 849-853. (1998).

[62]Risch, H. A., McLaughlin, J. R., Cole, D. E., et al."Prevalence and penetrance of germline BRCA1 and BRCA2 mutations in a population series of 649 women with ovarian cancer", Am J. Hum Genet, Vol. 68, pp. 700–710.( 2001).

[63]Roa, B. B., Boyd, A. A., Volcik, K., Richards, C. S. "Ashkenazi Jewish population frequencies for common mutations in BRCA1 and BRCA2",Nat Genet., Vol.14, pp. 185 –187. (1996).

[64]Seal, S., Thompson, D., Renwick,  A., et al. "Truncating mutations in the Fanconi anemia J gene BRIP1 are low-penetrance breast cancer susceptibility alleles", Nat Genet, Vol. 38, pp. 1239–1241. (2006).

[65]Shih, H. A., Couch, F. J., Nathanson, K. L., et al. "BRCA1 and BRCA2 mutation frequency in women evaluated in a breast cancer risk evaluation clinic", J. Clin. Oncol., Vol. 20, pp. 994–999. (2002).

[66]Simard, J., Tonin, P., Duroche,r F., et al. "Common origins of BRCA1 mutations in Canadian breast and ovarian cancer families" Nat Genet., Vol. 8, No. 4, pp. 392-398. (1994).

[67]Smith, S. A., Easton, D. F., Evans, D. G. and Ponder, B. A. "Allele losses in the region 17q12-21 in  familial breast and ovarian cancer involve the wild-type chromosome",Nat Genet, Vol. 2, pp. 128-131. (1992).

[68]Stanbridge, E. J. "Human tumor suppressor genes",Annu Rev Genet, Vol. 24, pp. 615-657. (1990).

[69]Struewing, J. P., Abeliovich, D., Peretz, T., Avishai, N., Kaback, M. M., Collins, F. S. and Brody, L. C."The carrier frequency of the BRCA1 185delAG mutation is approximately 1 percent in Ashkenazi Jewish individuals",Nat Genet, Vol. 11, pp. 198-200. (1995).

[70]Struewing, J. P., Hartge, P., Wacholder, S., et al. "The risk of cancer associated with specific mutations of BRCA1 and BRCA2 among Ashkenazi Jews", N Engl J Med., Vol. 336, No. 20, pp. 1401-1408. (1997).

[71]Szabo, C. I., King, M. C. "Population genetics of BRCA1 and BRCA2",  Am J. Hum Genet, Vol.  60, pp. 1013-1020. (1997).

[72]Tavtigian, S. V., Thomas, A., Frank, T. S. and Skolnick, M. H. "The BRCA1 gene and its protein product: characterization, therapeutic and diagnostic implications",AdvOncol, Vol. 14, pp. 3–13. (1998).

[73]Tikhomirova, L., Sinicka, O., Smite, D., Eglitis, J., Hodgson, S. V., Stengrevics, A. "High prevalence of two BRCA1 mutations, 4154delA and 5382insC, in Latvia",Fam Cancer., Vol.4, No.2, pp. 77-84. (2005).

[74]Trincado, P., Fardella, C., Mayerson, D., Montero, L., Obrien, A., Barrueto, K. and Gallegos, R."Prevalencia de la deleción 185AG del gen BRCA1   en mujereschilenas con cancer de mama", Rev Med Chile, Vol. 127, pp. 19-22. (1999).

[75]Vehmanen, P., Friedman, L. S., Eerola, H., et al. "Low proportion of BRCA1 and BRCA2 mutations in Finnish breast cancer families: evidence for additional susceptibility genes", Hum. Genet., Vol.6, No. 13, pp. 2309–2315. (1997).

[76]Warner, E., Foulkes, W., Goodwin, P., et al. "Preva-lence and penetrance of BRCA1 and BRCA2 gene mutations in unselected Ashkenazi Jewish women with breast cancer", J. Natl Cancer Inst, Vol. 91, pp. 1241-1247. (1999).

[77]Weitzel, J. N., Lagos, V., Blazer, K. R., Nelson, R., Ricker, C., Herzog, J., McGuire, C. and Neuhausen, S. "Prevalence of BRCA mutations and founder effect in high-risk Hispanic families; Cancer Epidemiol", Biomarkers Prev., Vol. 14, pp. 1666–1671. (2005).

[78]Wooster, R., Bignell, G., Lancaster, J., et al. "Identification of the breast cancer susceptibility gene BRCA2",Nature, Vol. 378, pp. 789 –92. (1995).

[79]Zhi, X., Szabo, C., Chopin, S., et al. "BRCA1 and BRCA2 sequence variants in Chinese breast cancer families", Hum. Mutat., Vol. 20, pp. 474. (2002).

Website

[1]Breast Cancer Information Core Database. [http://www.nhgri. nih.gov/Intramural_research/Lab_transfer/Bic/] (last access 15.09.2012). 

View Full Article