Male Breast Cancer and BRCA2: A Study from West Bengal, India

Male Breast Cancer and BRCA2: A Study from West Bengal, India | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Male Breast Cancer and BRCA2: A Study from West Bengal, India Abhishikta Ghosh Roy, Vanya Singh, Biswanath Sarkar, ARUP Ratan BANDYOPADHYAY This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5707242/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract One of the known risk factors for male breast cancer is germline mutations in the BRCA2 gene, which are linked to genomic alterations. Men carrying a germline BRCA2 mutation have a higher risk of developing breast carcinoma than the general population. Breast Cancer is the fifth leading cause of cancer-related deaths worldwide and the most common cancer in developed and developing countries. Male breast cancer accounts for approximately 1% of all breast cancer cases. The incidence of male breast cancer has increased over the past few decades, yet its etiology is still poorly understood, which may be due to the rarity of the disease and the lack of large-scale genetic epidemiologic studies. The present study aimed to evaluate the risk of developing Breast Carcinoma among Bengalee Hindu males. For the study, 36 male patients and 70 matched controls were included in this study. DNA sequencing and gene expression analysis revealed a significant association (p < 0.0001) between male breast cancer patients and BRCA2 mutations (rs1799943). There were many intronic variations whose associations were not determined. A 2.4-fold downregulation of tumour proteins was associated with patient risk, whereas controls showed a 1.28-fold upregulation. The present study, being the first attempt in Eastern India, envisaged a significant association (p < 0.001) of the disease for mutations in the exonic and flanking regions of the gene (OR-9.92; 95% CI = 4.40-22.37; p < 0.0001). Figures Figure 1 Figure 2 Figure 3 Figure 4 INTRODUCTION Male breast carcinoma accounts for less than 1% of all breast cancer cases in the United States (Landis et al., 1999 ; Lynch et al., 1999 ). Cancer of the male breast is uncommon (Giordano et al., 2002 ; Weiss et al., 2005 ) and accounts for approximately 1% of all breast cancer cases and 0.1% of cancer mortality in the male population in The USA (Weiss et al., 2005 ). Genetic research has revealed various distinctions between male and female breast cancer. For instance, mutations in BRCA1 play a minimal role in male breast cancer, whereas mutations in BRCA2 may be associated with up to 14% of male breast cancer cases. Male breast cancer is an uncommon disease, accounting for less than 1% of all breast cancers, and less than 1% of all cancers in men. The annual incidence is estimated to be approximately 1 per 100,000 men worldwide (Ly et al., 2013 ), with a lifetime risk of less than 1 in 1000. The identification of a small subset of individuals and families at increased risk of cancer due to mutations in cancer susceptibility genes is now possible with advances in molecular genetics (Timoteo et al., 2015). The incidence of male breast cancer increases with age, with the mean age at diagnosis ranging from 60 to 70 years (Korde et al., 2010 ). Understanding breast cancer in men is important because of its rarity. However, the management of male breast cancer is largely based on extrapolation of data from studies on female breast cancer (Cardoso et al., 2014; Soliman et al., 2014). Several unique characteristics of male breast cancer compared to female breast cancer have been identified, including a higher rate of hormone positivity, lower HER2 positivity, and more advanced presentation at diagnosis (Korde et al., 2015). The breast cancer susceptibility genes, BRCA1 and BRCA2, have large coding sequences of 51 exons, and many mutations have been characterized worldwide. Germline mutations in these genes together constitute approximately 6–7% of breast cancers, but the incidence and frequency vary between populations. Approximately 2,000 mutations have been reported in the Breast Cancer Information Core (BIC) database (Haiminen et al., 2010). Breast cancer has emerged as the most prevalent cancer in both developed and developing regions and is the fifth leading cause of cancer-related deaths worldwide, with 458,000 fatalities reported annually (Ferlay et al., 2010 ). Approximately 1% of breast cancer cases involve male breast cancer, which has increased in incidence over the past few decades, although its etiology remains unclear. Male breast cancer is a rare occurrence in the Western literature, accounting for less than 1% of all breast cancers (La Vecchia et al., 1992 ). Reports indicate that Indians present the disease at a more advanced stage, particularly at a younger age (Ihekwaba 1994 ; Jamal 2001 ; Vaizey et al. 1999 ). The risk factors for male breast cancer include a family history of the disease in first-degree female and male relatives, high hormone levels, particularly estrogen and prolactin, radiation exposure, hereditary factors, and conditions associated with hyperestrogenemia, such as cirrhosis of the liver or genetic syndromes, such as Klinefelter disease (D'Avanzo et al., 1995; Sasco et al., 1993 ; Koc et al., 2001). Although the epidemiological literature surrounding female breast cancer is extensive, male breast cancer remains poorly understood in terms of its epidemiology and etiology. Recent research suggests that male breast cancers may be a diverse group of malignancies that are molecularly and clinically distinct from those observed in women (Johansson et al., 2014). Female breast cancer is a complex and heterogeneous disease, with BRCA1 mutation carriers exhibiting specific pathological features including distinct morphology in female patients (Mavaddat et al., 2012). However, the pathological characteristics of breast cancers arising in male BRCA1/2 mutation carriers are not well understood, as the number of patients involved in each study was limited (Kwiatkowska et al., 2003; Ding et al., 2011; Ottini et al., 2012; Deb et al., 2012). Extensive knowledge of female BRCA1/2 and other inherited germline mutations is available, whereas little is known about male tumours. Therefore, the present study is a molecular anthropological attempt to gain insight into the BRCA2 status of Male Breast Cancer among the Bengalee Hindu Males of West Bengal. To the best of our knowledge, this is the first report of BRCA2 status in Male Breast Cancer patients in West Bengal. MATERIALS AND METHODS A retrospective investigation was carried out from April 2011 to March 2013, involving 36 male Bengalee Hindu patients with breast cancer. The study included histopathological confirmed breast carcinoma cases who attended cancer referral centers in Kolkata. Additionally, 70 controls matched for age, sex, and ethnic group were incorporated into the research. Detailed clinical information and family history were gathered for each patient through clinical examinations, with assistance from collaborating clinicians. The research received ethical approval from the relevant medical institutions, and all participants provided informed consent prior to enrolment in the study. For DNA isolation, approximately 5 mL of peripheral blood was collected from breast cancer patients and control subjects (individuals without any history of breast carcinoma) using BD Vacutainer K2 EDTA (6 mL) tubes, following written and informed consent. Genomic DNA was extracted from fresh whole blood utilizing the conventional phenol-chloroform method (Sambrook and Russel, 2001). The isolated genomic DNA was then dissolved in TE buffer (10 mM Tris-HCl and 0.1 mM EDTA, pH 8.0). PCR amplification was conducted to amplify exons and adjacent flanking regions. The reaction was performed in a total volume of 10.0 µL, containing 40–100 ng genomic DNA, 0.4 mM of each primer, 0.2 mM of each dNTP, 0.5–1.5 mM of MgCl2 (as appropriate), and 0.2 unit of Taq polymerase (Invitrogen, Carlsbad, CA) in a Thermocycler (GeneAmp-9700; PE Applied Biosystems, Foster City, CA). The annealing temperature was determined based on the Tm of the primer pairs. Exons 5–7 of the p53 gene were amplified using specifically designed primers (Table 1 ). The PCR-amplified DNA fragments were subsequently analyzed on 2% agarose gel and visualized through ethidium bromide staining. Mutation and Polymorphism Detection The PCR products, free of contaminating bands due to non-specific amplification, were directly sequenced forward and reverse in a DNA Analyzer 3730 (Applied Biosystems, USA). Nucleotide changes were detected by comparing the sequence obtained in the chromatogram with the normal p53 gene sequence using pair-wise BLAST (Tatusova et al., 1999) and SeqScape software v2.5 RESULTS Table 1 Age of the study cohort. Mean ± SD Range (Years) Total Patients 68.02 ± 0.383 42–82 36 Controls 69.65 ± 8.005 58–76 70 Table 2 Histopathological Characters and Treatment patterns of MBC: Characteristics % of Patients (n = 36) Age 50 77.77 Tumor Size 2cm 38.88 Lymph Node -ve 52.77 +ve 47.22 Estrogen Receptor -ve 27.77 +ve 72.22 Progesterone Receptor -ve 58.33 +ve 41.66 Operation Method BCS (Breast Conserving Surgery) 2.77 Masectomy 97.22 Chemotherapy Not Done 22.22 Done 77.77 Radiation Therapy Not Done 30.55 Done 69.44 Hormone Therapy Not Done 36.11 Done 63.88 Table 3 General Characteristics of Male Breast Cancer Patients. Characteristics % of Patients (n = 36) Chief Complaints Mass 57.14 Nipple Discharge 42.85 Duration of Symptoms Sym ≤ 6 months 32.14 6months 24months 21.42 Site of Tumor Right 37.5 Left 62.5 Location of Tumor Retroareolar 75.0 Other Quadrant 25.0 Table 1 illustrates the age distribution of the examined cohort. The average age of patients is 68.02 ± 0.383 years, whilst for controls it is 69.65 ± 8.005 years. Patient ages ranged from 42 to 82 years, and control ages spanned from 58 to 76 years. The study encompassed 36 histopathologically confirmed Male Breast Cancer (MBC) patients and 70 controls matched for age, sex, and ethnic group. Approximately 77.77% of patients were over 50 years old. Tumour dimensions varied, with the majority (61.11%) presenting tumours smaller than 2cm. Hormone-receptor analysis, conducted in collaboration with clinicians, revealed that 72.2% of patients were estrogen receptor-positive, while only 41.6% were progesterone receptor-positive. Despite recent technological advancements allowing breast preservation during tumour removal, nearly all patients (97.22%) chose mastectomy to prevent recurrence. Adjuvant treatments included chemotherapy for 77.77% of patients and radiotherapy for 69.4%. Daily hormone therapy with 20mg of tamoxifen was administered to 63.88% of patients. Male Breast Cancer symptoms ranged from asymptomatic to multiple manifestations. A mass or lump was reported by 57.14% of patients, while 42.85% experienced only nipple discharge. Notably, most patients (62.5%) had left breast tumours, and 75% presented with retroareolar tumours. Symptom duration varied between 6 and 12 months. Table 4 Disease Classification of the Patient Cohort, TNM Classification TNM Stage Percentage (%) (N = 36) T Stage T1 21.42 T2 14.28 T3 16.07 N Stage N1 7.14 N2 21.42 N3 14.28 M Stage M0 1.78 M1 3.57 Table 5 Mutation Spectrum of the Studied Cohort Exon/Intron Nucleotide Change Amino Acid Change Patient(%) N = 36 Control(%) N = 70 SNP Status P Value Exon 2 AGC > AAC Ser > Asp 16 (42.4) 2 (2.85) Reported (rs1799943) 0.0001 Exon 9 CCT > CTT Pro > Leu 9 (25.2) 3 (4.28) Reported (rs80359633) 0.0001 Intron 9 IVS9 + 139 T > C NA 7 (18.33) 1 (1.42) NOVEL 0.0001 Intron 9 IVS9 + 145 T > C NA 4 (14.07) 2 (2.85) NOVEL 0.0001 BRCA1 Mutations Patient (n = 36) Controls(n = 70) ODDS Ratio No. % No. % Present 29 80.55 8 11.43 34.0670*** 95%CI = 15.28–75.94 Absent 7 19.44 62 88.7 Total 36 100 70 100 BRCA2 (MIM: 600185; GenBank accession no. U43746) is a substantial gene that spans 70kb on genomic DNA at 13q12.13. It surpasses BRCA1 in size, comprising 27 exons, with 26 of them encoding a 3418 amino acid protein (Wooster et al., 1994; Tavtigian et al., 1996). A notable feature of BRCA2 is its expansive exon 11, which accounts for over 60% of the protein's encoding. Within exon 11, eight BRC repeats constitute the Rad51 binding domain (Pellegrini et al., 2002). The C-terminal domain, which is highly conserved (Takata et al., 2002), binds to ssDNA (Yang et al., 2002) and plays a crucial role in homologous recombination. In the population under investigation, exons 2, 8, 9, 13, and 22 of the BRCA2 gene underwent sequencing. The molecular analysis revealed three intronic mutations and two exonic mutations. The intronic variant does not alter amino acids. A significant association (p < 0.0001) was found with the exon two mutation (rs1799943), present in 42.4% of patients and 2.85% of controls, resulting in an amino acid change from Serine to Asparagine. Additionally, a mutation in the coding region of exon 9 (rs80359633) changes proline to leucine in 25.2% of patients and 4.28% of controls. Intron 9 (rs2126042) mutations were identified in 18.33% of patients and 1.42% of controls. Two novel variants were discovered in Intron 9: IVS9 + 139 T > C and IVS9 + 145 T > C, occurring in 14.7% of patients and 2.85% of controls (Figs. 1 and 2 ). Regarding gene expression, BRCA is expressed at low levels across various normal and pathological human tissues, indicating a general role in cellular homeostasis. The BRCA tumour suppressor gene encodes a 220-kDa nuclear phosphoprotein that contributes to the DNA damage response and maintains genomic integrity. In addition,, BRCA has been found to regulate gene transcription. The broad caretaker function of BRCA does not explain why mutations lead to cancer only in certain organs, suggesting the involvement of tissue-specific coregulators. Therefore, it is hypothesized that dysregulation of BRCA gene expression in human samples, especially in sporadic breast cancer, might also account for pathogenesis. To determine the putative variations in BRCA expression levels, RT-PCR was performed to amplify a chimeric BRCA synthetic transcript and the endogenous messenger RNA. Following RNA extraction from tissue samples, gene expression analysis was conducted. This involved converting the BRCA gene to cDNA and subsequently performing a gene expression study using Real-Time PCR (RT-PCR, ABI 7500). The results were analysed through log transformation of the ct value. As BRCA functions as a tumour suppressor gene, breast cancer patients exhibited reduced BRCA protein levels, leading to impaired tumour suppression and disease progression. The findings revealed a 2.4-fold decrease in BRCA protein expression in patients, compared to a 1.28-fold regulation observed in control subjects (Figs. 4 and 4). DISCUSSION Chromosome 13q12.13 houses the BRCA2 gene, which has been linked to the majority of hereditary breast cancer cases in males (Wooster et al., 1994). The BRCA2 gene, which is substantial in size, has recently been found to regulate intracellular localisation and DNA binding capacity. The loss of these functions, resulting from BRCA2 inactivation, may be a crucial factor in genomic instability and tumour formation (Davies et al., 2001). BRCA2 genes are categorised as tumour suppressors, maintaining genomic stability and cell cycle checkpoint control (Ottini et al., 2008). Mutations and instabilities in BRCA2 can ultimately lead to cancer development. The reported prevalence of BRCA2 germline mutations in Male Breast Cancer varies across populations, likely due to limited sample sizes, diverse methodologies, and differing sensitivities of mutation detection techniques. BRCA1 mutations are uncommon in Male Breast Cancer and are estimated to occur in over 5% of all female Breast Cancers. Numerous studies have sought to elucidate the role of BRCA1 mutations. The majority found no carriers of BRCA1 mutations, although a few investigations have identified BRCA1 mutations in MBC (Sverdlov et al., 2000; Basham et al., 2002; Csokay et al., 1999; Tirkkonen et al., 1999). This study represents the first known examination of the genetic landscape of Male Breast Cancer in West Bengal, detailing the genotypic, conventional clinicopathological, and intrinsic phenotypic characteristics of this condition. Male breast cancer constitutes less than 1% of all Breast Cancer cases, with limited understanding of its epidemiological, genetic, and clinicopathological aspects. The research revealed that breast cancer in male BRCA2 mutation carriers exhibited significantly higher stage and histologic grade, and more frequently displayed ER + and PR + status. Men are more likely to be diagnosed with advanced-stage breast cancer compared to women (Giardano et al., 2004). This disparity is generally attributed to delayed diagnosis in a population unaware of their risk and not routinely screened for breast cancer. Although male breast cancer primaries tend to be slightly smaller at initial diagnosis, they more often present with locoregional metastasis. Notably, this study found that 52.77% of male BRCA2 mutation carriers presented with lymph node involvement. The research involved sequencing BRCA2 exons 2, 8, 9 and 13. Previous studies have shown that exonic variants can affect pre-mRNA splicing by altering splice sites or modifying splicing regulatory elements (Di Giacomo et al., 2013). The analysis identified four sequence variants, two of which are novel. A previous study from Southern India reported a novel variant in exon 8 with a T-C transition in female breast cancer. T and C transitions were also common in this study, albeit with changes at different gene bases. Three intronic variants of unknown significance were identified in Intron 9. The sequence variant in exon 9 results in an amino acid change from Proline to Leucine. It is worth noting that BRCA2 mutations are generally less prevalent than BRCA1 mutations. The gene expression study (Fig. 24, 26) using Real-Time PCR (RT-PCR) indicated a reduction in tumour protein expression of both p53 and BRCA genes within the examined Bengalee population. Nevertheless, numerous studies have noted variations in SNP allele frequencies across ethnic groups (Goddard et al. 2000; Excoffier and Slatkin 1995; Bonnen et al. 2000; Tishkoff et al. 2000; Niu et al. 2002). Consequently, SNP variations and decreased gene expression are not the sole factors contributing to breast cancer. In male breast cancer (MBC), lymph node involvement is a crucial predictor, although there is a shortage of comprehensive research on adjuvant radiotherapy. Several studies suggest that MBC frequently involves the nipple and skin. A notably high rate of hormone receptor expression is among the prognostic factors for MBC. Historically, MBC was thought to have a worse prognosis than its female counterpart, with lower survival rates attributed to older age and advanced disease at diagnosis. Comparative analyses show that men generally have poorer survival rates than women. Study outcomes: This pioneering study from India revealed the spectrum and prevalence of the BRCA2 gene in the studied population. The research also demonstrated that a mutation in the tumour suppressor gene alone is insufficient to trigger an individual's lifetime risk of developing the disease. Genetic screening for mutations and environmental factor modifications could potentially be utilised for disease prognosis and benefit overall public health in men. Declarations We, the undersigned authors, certify that this manuscript is an original work and has not been submitted or published elsewhere. All authors have made significant contributions to the conception, design, analysis, and/or interpretation of the research, and have read and approved the final manuscript. The authors declare no conflicts of interest regarding the publication of this manuscript. This work was supported by Anthropological Survey of India, Ministry of Culture, Government of India. The funders had no role in the design, data collection, analysis, decision to publish, or preparation of the manuscript. This study was conducted in accordance with ethical standards and guidelines. Approval was obtained from Institutional Ethical Committee of Anthropological Survey of India and collaborators. Written informed consent was obtained from all participants involved in the study. We acknowledge all the study participants for the study. Author Contribution Equal contribution of all authors References Ly D, Forman D, Ferlay J, Brinton LA, Cook MB. An International comparison of male and female breast cancer incident rates. Intl J Cancer. 2013; 132 (8): 1918-26. Korde LA, Zujewski JA,Kamin L, Giordano S, Domchek S, Anderson WF et al.,. Multidisciplinary meeting on male breast cancer : Summary and research recommendations. J Clin Oncol. 2010; 28 (12) : 2114 – 22. La Vecchia C, Levi F, Lucchini F (1992). Descriptive epidemiology of male breast cancer in Europe. Int J Cancer, 51, 62-6. Ihekwaba FN (1994). Breast cancer in men in Black Africa: a report of 73 cases. J R Coll Surg Edinb , 39 , 344-7. Jamal AA (2001). Pattern of breast diseases in a teaching hospitalin Jeddah, Saudi Arabia. Saudi Med J , 22 , 110-13. Vaizey C, Burke M, Lange M (1999). Carcinoma of the male breast– a review of 91 patients from the Johannesburg Hospital breast clinics. S Afr J Surg , 37 , 6-8. D’ Avanzo B, La Vecchia C (1995). Risk factors for male breastcancer. Br J Cancer , 71 , 1359-62. Sasco AJ, Lowenfels AB, Pasker-de Jong P (1993). Epidemiology of male breast cancer. A meta-analysis of published case-control studies and discussion of selected aetiological factors. Int J Cancer , 53, 538-49. Koc M, Polat P (2001). Epidemiology and aetiological factors of male breast cancer: a ten years retrospective study in eastern Turkey. Eur J Cancer Prev , 10 , 531-4. Landis SH, Murray T, Bolden S, Wingo PA. Cancer statistics, 1999. CA Cancer J Clin 1999;49:8–31.[PubMed: 10200775] Lynch HT, Watson P, Narod SA. The genetic epidemiology of male breast carcinoma. Cancer 1999;86:744–746. [PubMed: 10463967] Ferlay J, Shin HR, Bray F, et al (2010). Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer , 127 , 2893-917. Giordano SH, Buzdar AU, Hortobagyi GN (2002). Breast cancer in men. Ann Intern Med , 137 , 678-87. Weiss JR, Moysich KB, Swede H (2005). Epidemiology of male breast cancer. Cancer Epidemiol Biomarkers Prev, 14, 20-6. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5707242","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":398332614,"identity":"27e27d8a-ff71-4fe7-9633-702b94bf1d9e","order_by":0,"name":"Abhishikta Ghosh 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15:53:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5707242/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5707242/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":73269504,"identity":"697c68dd-92ce-4bea-8046-8403326e3b0f","added_by":"auto","created_at":"2025-01-08 10:44:27","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":229283,"visible":true,"origin":"","legend":"\u003cp\u003eGENE SEQUENCING RESULTS\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5707242/v1/bdc3b02389467bb0f770b95f.png"},{"id":73269502,"identity":"66cabb1f-969e-4127-87ec-9d43ed0a6b88","added_by":"auto","created_at":"2025-01-08 10:44:27","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":10137,"visible":true,"origin":"","legend":"\u003cp\u003ePatient Control Graph of Mutation 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RT-PCR\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5707242/v1/a2a2bccc77a5af282a37c9e4.png"},{"id":86484982,"identity":"2957ed5d-e442-468a-b5c4-7e997239813a","added_by":"auto","created_at":"2025-07-11 08:09:06","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":995250,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5707242/v1/aa6aa5d2-7cef-4fcf-bbc3-22f1071772dd.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eMale Breast Cancer and BRCA2: A Study from West Bengal, India\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eMale breast carcinoma accounts for less than 1% of all breast cancer cases in the United States (Landis et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Lynch et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). Cancer of the male breast is uncommon (Giordano et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Weiss et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) and accounts for approximately 1% of all breast cancer cases and 0.1% of cancer mortality in the male population in The USA (Weiss et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2005\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGenetic research has revealed various distinctions between male and female breast cancer. For instance, mutations in BRCA1 play a minimal role in male breast cancer, whereas mutations in BRCA2 may be associated with up to 14% of male breast cancer cases. Male breast cancer is an uncommon disease, accounting for less than 1% of all breast cancers, and less than 1% of all cancers in men. The annual incidence is estimated to be approximately 1 per 100,000 men worldwide (Ly et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), with a lifetime risk of less than 1 in 1000. The identification of a small subset of individuals and families at increased risk of cancer due to mutations in cancer susceptibility genes is now possible with advances in molecular genetics (Timoteo et al., 2015). The incidence of male breast cancer increases with age, with the mean age at diagnosis ranging from 60 to 70 years (Korde et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Understanding breast cancer in men is important because of its rarity.\u003c/p\u003e \u003cp\u003eHowever, the management of male breast cancer is largely based on extrapolation of data from studies on female breast cancer (Cardoso et al., 2014; Soliman et al., 2014). Several unique characteristics of male breast cancer compared to female breast cancer have been identified, including a higher rate of hormone positivity, lower HER2 positivity, and more advanced presentation at diagnosis (Korde et al., 2015). The breast cancer susceptibility genes, BRCA1 and BRCA2, have large coding sequences of 51 exons, and many mutations have been characterized worldwide. Germline mutations in these genes together constitute approximately 6\u0026ndash;7% of breast cancers, but the incidence and frequency vary between populations. Approximately 2,000 mutations have been reported in the Breast Cancer Information Core (BIC) database (Haiminen et al., 2010).\u003c/p\u003e \u003cp\u003eBreast cancer has emerged as the most prevalent cancer in both developed and developing regions and is the fifth leading cause of cancer-related deaths worldwide, with 458,000 fatalities reported annually (Ferlay et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Approximately 1% of breast cancer cases involve male breast cancer, which has increased in incidence over the past few decades, although its etiology remains unclear. Male breast cancer is a rare occurrence in the Western literature, accounting for less than 1% of all breast cancers (La Vecchia et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Reports indicate that Indians present the disease at a more advanced stage, particularly at a younger age (Ihekwaba \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Jamal \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Vaizey et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). The risk factors for male breast cancer include a family history of the disease in first-degree female and male relatives, high hormone levels, particularly estrogen and prolactin, radiation exposure, hereditary factors, and conditions associated with hyperestrogenemia, such as cirrhosis of the liver or genetic syndromes, such as Klinefelter disease (D'Avanzo et al., 1995; Sasco et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Koc et al., 2001). Although the epidemiological literature surrounding female breast cancer is extensive, male breast cancer remains poorly understood in terms of its epidemiology and etiology.\u003c/p\u003e \u003cp\u003eRecent research suggests that male breast cancers may be a diverse group of malignancies that are molecularly and clinically distinct from those observed in women (Johansson et al., 2014). Female breast cancer is a complex and heterogeneous disease, with BRCA1 mutation carriers exhibiting specific pathological features including distinct morphology in female patients (Mavaddat et al., 2012). However, the pathological characteristics of breast cancers arising in male BRCA1/2 mutation carriers are not well understood, as the number of patients involved in each study was limited (Kwiatkowska et al., 2003; Ding et al., 2011; Ottini et al., 2012; Deb et al., 2012). Extensive knowledge of female BRCA1/2 and other inherited germline mutations is available, whereas little is known about male tumours. Therefore, the present study is a molecular anthropological attempt to gain insight into the BRCA2 status of Male Breast Cancer among the Bengalee Hindu Males of West Bengal. To the best of our knowledge, this is the first report of BRCA2 status in Male Breast Cancer patients in West Bengal.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eA retrospective investigation was carried out from April 2011 to March 2013, involving 36 male Bengalee Hindu patients with breast cancer. The study included histopathological confirmed breast carcinoma cases who attended cancer referral centers in Kolkata. Additionally, 70 controls matched for age, sex, and ethnic group were incorporated into the research. Detailed clinical information and family history were gathered for each patient through clinical examinations, with assistance from collaborating clinicians.\u003c/p\u003e \u003cp\u003eThe research received ethical approval from the relevant medical institutions, and all participants provided informed consent prior to enrolment in the study.\u003c/p\u003e \u003cp\u003eFor DNA isolation, approximately 5 mL of peripheral blood was collected from breast cancer patients and control subjects (individuals without any history of breast carcinoma) using BD Vacutainer K2 EDTA (6 mL) tubes, following written and informed consent. Genomic DNA was extracted from fresh whole blood utilizing the conventional phenol-chloroform method (Sambrook and Russel, 2001). The isolated genomic DNA was then dissolved in TE buffer (10 mM Tris-HCl and 0.1 mM EDTA, pH 8.0).\u003c/p\u003e \u003cp\u003ePCR amplification was conducted to amplify exons and adjacent flanking regions. The reaction was performed in a total volume of 10.0 \u0026micro;L, containing 40\u0026ndash;100 ng genomic DNA, 0.4 mM of each primer, 0.2 mM of each dNTP, 0.5\u0026ndash;1.5 mM of MgCl2 (as appropriate), and 0.2 unit of Taq polymerase (Invitrogen, Carlsbad, CA) in a Thermocycler (GeneAmp-9700; PE Applied Biosystems, Foster City, CA). The annealing temperature was determined based on the Tm of the primer pairs. Exons 5\u0026ndash;7 of the p53 gene were amplified using specifically designed primers (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The PCR-amplified DNA fragments were subsequently analyzed on 2% agarose gel and visualized through ethidium bromide staining.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMutation and Polymorphism Detection\u003c/h2\u003e \u003cp\u003eThe PCR products, free of contaminating bands due to non-specific amplification, were directly sequenced forward and reverse in a DNA Analyzer 3730 (Applied Biosystems, USA). Nucleotide changes were detected by comparing the sequence obtained in the chromatogram with the normal p53 gene sequence using pair-wise BLAST (Tatusova et al., 1999) and SeqScape software v2.5\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAge of the study cohort.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRange (Years)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePatients\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e68.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.383\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42\u0026ndash;82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControls\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e69.65\u0026thinsp;\u0026plusmn;\u0026thinsp;8.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58\u0026ndash;76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eHistopathological Characters and Treatment patterns of MBC:\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eCharacteristics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e% of Patients (n\u0026thinsp;=\u0026thinsp;36)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAge\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e77.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eTumor Size\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;2cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e61.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;2cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e38.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLymph Node\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-ve\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e52.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+ve\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eEstrogen Receptor\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-ve\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e27.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+ve\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e72.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eProgesterone Receptor\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-ve\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+ve\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e41.66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eOperation Method\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBCS (Breast Conserving Surgery)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMasectomy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e97.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eChemotherapy\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNot Done\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e77.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eRadiation Therapy\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNot Done\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e69.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eHormone Therapy\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNot Done\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e36.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e63.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eGeneral Characteristics of Male Breast Cancer Patients.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eCharacteristics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e% of Patients (n\u0026thinsp;=\u0026thinsp;36)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eChief Complaints\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMass\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNipple Discharge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eDuration of Symptoms\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSym\u0026thinsp;\u0026le;\u0026thinsp;6 months\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6months\u0026thinsp;\u0026lt;\u0026thinsp;Sym\u0026thinsp;\u0026le;\u0026thinsp;12months\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e39.29\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12months\u0026thinsp;\u0026le;\u0026thinsp;Sym\u0026thinsp;\u0026le;\u0026thinsp;24months\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSym\u0026thinsp;\u0026gt;\u0026thinsp;24months\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSite of Tumor\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLeft\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLocation of Tumor\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRetroareolar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOther Quadrant\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e illustrates the age distribution of the examined cohort. The average age of patients is 68.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.383 years, whilst for controls it is 69.65\u0026thinsp;\u0026plusmn;\u0026thinsp;8.005 years. Patient ages ranged from 42 to 82 years, and control ages spanned from 58 to 76 years. The study encompassed 36 histopathologically confirmed Male Breast Cancer (MBC) patients and 70 controls matched for age, sex, and ethnic group. Approximately 77.77% of patients were over 50 years old. Tumour dimensions varied, with the majority (61.11%) presenting tumours smaller than 2cm. Hormone-receptor analysis, conducted in collaboration with clinicians, revealed that 72.2% of patients were estrogen receptor-positive, while only 41.6% were progesterone receptor-positive. Despite recent technological advancements allowing breast preservation during tumour removal, nearly all patients (97.22%) chose mastectomy to prevent recurrence. Adjuvant treatments included chemotherapy for 77.77% of patients and radiotherapy for 69.4%. Daily hormone therapy with 20mg of tamoxifen was administered to 63.88% of patients.\u003c/p\u003e \u003cp\u003eMale Breast Cancer symptoms ranged from asymptomatic to multiple manifestations. A mass or lump was reported by 57.14% of patients, while 42.85% experienced only nipple discharge. Notably, most patients (62.5%) had left breast tumours, and 75% presented with retroareolar tumours. Symptom duration varied between 6 and 12 months.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDisease Classification of the Patient Cohort, TNM Classification\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNM Stage\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePercentage (%) (N\u0026thinsp;=\u0026thinsp;36)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eT Stage\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16.07\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eN Stage\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eM Stage\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eM0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.78\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eM1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMutation Spectrum of the Studied Cohort\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExon/Intron\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNucleotide Change\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAmino Acid Change\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePatient(%)\u003c/p\u003e \u003cp\u003eN\u0026thinsp;=\u0026thinsp;36\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl(%)\u003c/p\u003e \u003cp\u003eN\u0026thinsp;=\u0026thinsp;70\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSNP Status\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eP Value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExon 2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAGC\u0026thinsp;\u0026gt;\u0026thinsp;AAC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSer\u0026thinsp;\u0026gt;\u0026thinsp;Asp\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e16 (42.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2 (2.85)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eReported\u003c/p\u003e \u003cp\u003e(rs1799943)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.0001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExon 9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCCT\u0026thinsp;\u0026gt;\u0026thinsp;CTT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePro\u0026thinsp;\u0026gt;\u0026thinsp;Leu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9 (25.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3 (4.28)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eReported\u003c/p\u003e \u003cp\u003e(rs80359633)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.0001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntron 9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIVS9\u0026thinsp;+\u0026thinsp;139 T\u0026thinsp;\u0026gt;\u0026thinsp;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7 (18.33)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1 (1.42)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNOVEL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.0001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntron 9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIVS9\u0026thinsp;+\u0026thinsp;145 T\u0026thinsp;\u0026gt;\u0026thinsp;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4 (14.07)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2 (2.85)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNOVEL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.0001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eBRCA1 Mutations\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003ePatient (n\u0026thinsp;=\u0026thinsp;36)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eControls(n\u0026thinsp;=\u0026thinsp;70)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eODDS Ratio\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePresent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e80.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003e34.0670***\u003c/p\u003e \u003cp\u003e95%CI\u0026thinsp;=\u0026thinsp;15.28\u0026ndash;75.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAbsent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e88.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eBRCA2 (MIM: 600185; GenBank accession no. U43746) is a substantial gene that spans 70kb on genomic DNA at 13q12.13. It surpasses BRCA1 in size, comprising 27 exons, with 26 of them encoding a 3418 amino acid protein (Wooster et al., 1994; Tavtigian et al., 1996). A notable feature of BRCA2 is its expansive exon 11, which accounts for over 60% of the protein's encoding. Within exon 11, eight BRC repeats constitute the Rad51 binding domain (Pellegrini et al., 2002). The C-terminal domain, which is highly conserved (Takata et al., 2002), binds to ssDNA (Yang et al., 2002) and plays a crucial role in homologous recombination.\u003c/p\u003e \u003cp\u003eIn the population under investigation, exons 2, 8, 9, 13, and 22 of the BRCA2 gene underwent sequencing. The molecular analysis revealed three intronic mutations and two exonic mutations. The intronic variant does not alter amino acids. A significant association (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) was found with the exon two mutation (rs1799943), present in 42.4% of patients and 2.85% of controls, resulting in an amino acid change from Serine to Asparagine. Additionally, a mutation in the coding region of exon 9 (rs80359633) changes proline to leucine in 25.2% of patients and 4.28% of controls. Intron 9 (rs2126042) mutations were identified in 18.33% of patients and 1.42% of controls. Two novel variants were discovered in Intron 9: IVS9\u0026thinsp;+\u0026thinsp;139 T\u0026thinsp;\u0026gt;\u0026thinsp;C and IVS9\u0026thinsp;+\u0026thinsp;145 T\u0026thinsp;\u0026gt;\u0026thinsp;C, occurring in 14.7% of patients and 2.85% of controls (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eRegarding gene expression, BRCA is expressed at low levels across various normal and pathological human tissues, indicating a general role in cellular homeostasis. The BRCA tumour suppressor gene encodes a 220-kDa nuclear phosphoprotein that contributes to the DNA damage response and maintains genomic integrity. In addition,, BRCA has been found to regulate gene transcription. The broad caretaker function of BRCA does not explain why mutations lead to cancer only in certain organs, suggesting the involvement of tissue-specific coregulators. Therefore, it is hypothesized that dysregulation of BRCA gene expression in human samples, especially in sporadic breast cancer, might also account for pathogenesis. To determine the putative variations in BRCA expression levels, RT-PCR was performed to amplify a chimeric BRCA synthetic transcript and the endogenous messenger RNA.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFollowing RNA extraction from tissue samples, gene expression analysis was conducted. This involved converting the BRCA gene to cDNA and subsequently performing a gene expression study using Real-Time PCR (RT-PCR, ABI 7500). The results were analysed through log transformation of the ct value. As BRCA functions as a tumour suppressor gene, breast cancer patients exhibited reduced BRCA protein levels, leading to impaired tumour suppression and disease progression. The findings revealed a 2.4-fold decrease in BRCA protein expression in patients, compared to a 1.28-fold regulation observed in control subjects (Figs.\u0026nbsp;4 and 4).\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eChromosome 13q12.13 houses the BRCA2 gene, which has been linked to the majority of hereditary breast cancer cases in males (Wooster et al., 1994). The BRCA2 gene, which is substantial in size, has recently been found to regulate intracellular localisation and DNA binding capacity. The loss of these functions, resulting from BRCA2 inactivation, may be a crucial factor in genomic instability and tumour formation (Davies et al., 2001). BRCA2 genes are categorised as tumour suppressors, maintaining genomic stability and cell cycle checkpoint control (Ottini et al., 2008). Mutations and instabilities in BRCA2 can ultimately lead to cancer development. The reported prevalence of BRCA2 germline mutations in Male Breast Cancer varies across populations, likely due to limited sample sizes, diverse methodologies, and differing sensitivities of mutation detection techniques. BRCA1 mutations are uncommon in Male Breast Cancer and are estimated to occur in over 5% of all female Breast Cancers. Numerous studies have sought to elucidate the role of BRCA1 mutations. The majority found no carriers of BRCA1 mutations, although a few investigations have identified BRCA1 mutations in MBC (Sverdlov et al., 2000; Basham et al., 2002; Csokay et al., 1999; Tirkkonen et al., 1999).\u003c/p\u003e \u003cp\u003eThis study represents the first known examination of the genetic landscape of Male Breast Cancer in West Bengal, detailing the genotypic, conventional clinicopathological, and intrinsic phenotypic characteristics of this condition. Male breast cancer constitutes less than 1% of all Breast Cancer cases, with limited understanding of its epidemiological, genetic, and clinicopathological aspects.\u003c/p\u003e \u003cp\u003eThe research revealed that breast cancer in male BRCA2 mutation carriers exhibited significantly higher stage and histologic grade, and more frequently displayed ER\u0026thinsp;+\u0026thinsp;and PR\u0026thinsp;+\u0026thinsp;status. Men are more likely to be diagnosed with advanced-stage breast cancer compared to women (Giardano et al., 2004). This disparity is generally attributed to delayed diagnosis in a population unaware of their risk and not routinely screened for breast cancer. Although male breast cancer primaries tend to be slightly smaller at initial diagnosis, they more often present with locoregional metastasis. Notably, this study found that 52.77% of male BRCA2 mutation carriers presented with lymph node involvement.\u003c/p\u003e \u003cp\u003eThe research involved sequencing BRCA2 exons 2, 8, 9 and 13. Previous studies have shown that exonic variants can affect pre-mRNA splicing by altering splice sites or modifying splicing regulatory elements (Di Giacomo et al., 2013). The analysis identified four sequence variants, two of which are novel. A previous study from Southern India reported a novel variant in exon 8 with a T-C transition in female breast cancer. T and C transitions were also common in this study, albeit with changes at different gene bases. Three intronic variants of unknown significance were identified in Intron 9. The sequence variant in exon 9 results in an amino acid change from Proline to Leucine. It is worth noting that BRCA2 mutations are generally less prevalent than BRCA1 mutations.\u003c/p\u003e \u003cp\u003eThe gene expression study (Fig.\u0026nbsp;24, 26) using Real-Time PCR (RT-PCR) indicated a reduction in tumour protein expression of both p53 and BRCA genes within the examined Bengalee population. Nevertheless, numerous studies have noted variations in SNP allele frequencies across ethnic groups (Goddard et al. 2000; Excoffier and Slatkin 1995; Bonnen et al. 2000; Tishkoff et al. 2000; Niu et al. 2002). Consequently, SNP variations and decreased gene expression are not the sole factors contributing to breast cancer.\u003c/p\u003e \u003cp\u003eIn male breast cancer (MBC), lymph node involvement is a crucial predictor, although there is a shortage of comprehensive research on adjuvant radiotherapy. Several studies suggest that MBC frequently involves the nipple and skin. A notably high rate of hormone receptor expression is among the prognostic factors for MBC.\u003c/p\u003e \u003cp\u003eHistorically, MBC was thought to have a worse prognosis than its female counterpart, with lower survival rates attributed to older age and advanced disease at diagnosis. Comparative analyses show that men generally have poorer survival rates than women.\u003c/p\u003e\n\u003ch3\u003eStudy outcomes:\u003c/h3\u003e\n\u003cp\u003eThis pioneering study from India revealed the spectrum and prevalence of the BRCA2 gene in the studied population. The research also demonstrated that a mutation in the tumour suppressor gene alone is insufficient to trigger an individual's lifetime risk of developing the disease. Genetic screening for mutations and environmental factor modifications could potentially be utilised for disease prognosis and benefit overall public health in men.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eWe, the undersigned authors, certify that this manuscript is an original work and has not been submitted or published elsewhere. All authors have made significant contributions to the conception, design, analysis, and/or interpretation of the research, and have read and approved the final manuscript. The authors declare no conflicts of interest regarding the publication of this manuscript. This work was supported by Anthropological Survey of India, Ministry of Culture, Government of India. The funders had no role in the design, data collection, analysis, decision to publish, or preparation of the manuscript. This study was conducted in accordance with ethical standards and guidelines. Approval was obtained from Institutional Ethical Committee of Anthropological Survey of India and collaborators. Written informed consent was obtained from all participants involved in the study. We acknowledge all the study participants for the study.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eEqual contribution of all authors\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eLy D, Forman D, Ferlay J, Brinton LA, Cook MB. An International comparison of male and female breast cancer incident rates. Intl J Cancer. 2013; 132 (8): 1918-26.\u003c/li\u003e\n\u003cli\u003eKorde LA, Zujewski JA,Kamin L, Giordano S, Domchek S, Anderson WF et al.,. Multidisciplinary meeting on male breast cancer : Summary and research recommendations. J Clin Oncol. 2010; 28 (12) : 2114 \u0026ndash; 22. \u003c/li\u003e\n\u003cli\u003eLa Vecchia C, Levi F, Lucchini F (1992). Descriptive epidemiology of male breast cancer in Europe. Int J Cancer, 51, 62-6.\u003c/li\u003e\n\u003cli\u003eIhekwaba FN (1994). Breast cancer in men in Black Africa: a report of 73 cases. \u003cem\u003eJ R Coll Surg Edinb\u003c/em\u003e, \u003cstrong\u003e39\u003c/strong\u003e, 344-7. \u003c/li\u003e\n\u003cli\u003eJamal AA (2001). Pattern of breast diseases in a teaching hospitalin Jeddah, Saudi Arabia. \u003cem\u003eSaudi Med J\u003c/em\u003e, \u003cstrong\u003e22\u003c/strong\u003e, 110-13.\u003c/li\u003e\n\u003cli\u003eVaizey C, Burke M, Lange M (1999). Carcinoma of the male breast\u0026ndash; a review of 91 patients from the Johannesburg Hospital breast clinics. \u003cem\u003eS Afr J Surg\u003c/em\u003e, \u003cstrong\u003e37\u003c/strong\u003e, 6-8.\u003c/li\u003e\n\u003cli\u003eD\u0026rsquo; Avanzo B, La Vecchia C (1995). Risk factors for male breastcancer. \u003cem\u003eBr J Cancer\u003c/em\u003e, \u003cstrong\u003e71\u003c/strong\u003e, 1359-62.\u003c/li\u003e\n\u003cli\u003eSasco AJ, Lowenfels AB, Pasker-de Jong P (1993). Epidemiology of male breast cancer. A meta-analysis of published case-control studies and discussion of selected aetiological factors. \u003cem\u003eInt J \u003c/em\u003e\u003cem\u003eCancer\u003c/em\u003e, 53, 538-49.\u003c/li\u003e\n\u003cli\u003eKoc M, Polat P (2001). Epidemiology and aetiological factors of male breast cancer: a ten years retrospective study in eastern Turkey. \u003cem\u003eEur J Cancer Prev\u003c/em\u003e, \u003cstrong\u003e10\u003c/strong\u003e, 531-4.\u003c/li\u003e\n\u003cli\u003eLandis SH, Murray T, Bolden S, Wingo PA. Cancer statistics, 1999. CA Cancer J Clin 1999;49:8\u0026ndash;31.[PubMed: 10200775]\u003c/li\u003e\n\u003cli\u003eLynch HT, Watson P, Narod SA. The genetic epidemiology of male breast carcinoma. Cancer 1999;86:744\u0026ndash;746. [PubMed: 10463967]\u003c/li\u003e\n\u003cli\u003eFerlay J, Shin HR, Bray F, et al (2010). Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. \u003cem\u003eInt J Cancer\u003c/em\u003e, \u003cstrong\u003e127\u003c/strong\u003e, 2893-917.\u003c/li\u003e\n\u003cli\u003eGiordano SH, Buzdar AU, Hortobagyi GN (2002). Breast cancer in men. \u003cem\u003eAnn Intern Med\u003c/em\u003e, \u003cstrong\u003e137\u003c/strong\u003e, 678-87.\u003c/li\u003e\n\u003cli\u003eWeiss JR, Moysich KB, Swede H (2005). Epidemiology of male breast cancer. Cancer Epidemiol Biomarkers Prev, 14, 20-6.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-5707242/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5707242/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOne of the known risk factors for male breast cancer is germline mutations in the BRCA2 gene, which are linked to genomic alterations. Men carrying a germline BRCA2 mutation have a higher risk of developing breast carcinoma than the general population. Breast Cancer is the fifth leading cause of cancer-related deaths worldwide and the most common cancer in developed and developing countries. Male breast cancer accounts for approximately 1% of all breast cancer cases. The incidence of male breast cancer has increased over the past few decades, yet its etiology is still poorly understood, which may be due to the rarity of the disease and the lack of large-scale genetic epidemiologic studies. The present study aimed to evaluate the risk of developing Breast Carcinoma among Bengalee Hindu males. For the study, 36 male patients and 70 matched controls were included in this study. DNA sequencing and gene expression analysis revealed a significant association (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) between male breast cancer patients and BRCA2 mutations (rs1799943). There were many intronic variations whose associations were not determined. A 2.4-fold downregulation of tumour proteins was associated with patient risk, whereas controls showed a 1.28-fold upregulation. The present study, being the first attempt in Eastern India, envisaged a significant association (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) of the disease for mutations in the exonic and flanking regions of the gene (OR-9.92; 95% CI\u0026thinsp;=\u0026thinsp;4.40-22.37; p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001).\u003c/p\u003e","manuscriptTitle":"Male Breast Cancer and BRCA2: A Study from West Bengal, India","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-08 10:44:22","doi":"10.21203/rs.3.rs-5707242/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a8f025b7-ed40-4565-8390-cd1d67adfd6a","owner":[],"postedDate":"January 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-07-11T08:08:50+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-08 10:44:22","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5707242","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5707242","identity":"rs-5707242","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}