Epidemiology of Breast Cancer among Male in University of Abuja Teaching Hospital Gwagwalada

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Dangana .A,

Hematology Laboratory, University of Abuja Teaching Hospital, Abuja
Ishaku .H, Victoria .I,

Histology Laboratory, University of Abuja Teaching Hospital, Abuja
Christy .C.F,

College of Health Sciences, Pathology Department, University of Abuja
Egenti B.N

College of Health Sciences, Community Medicine Department, University of Abuja

All correspondence to: isalemit@yahoo.co.uk

ABSTRACT
Breast Cancer is the most common non-communicable disease and non-cutaneous malignancy. it’s a rare disease among men which account for less than 1% of all cancers in men. We examined the trends in the prevalence rate of breast cancer in men among tissues submitted to histopathology laboratory university of Abuja Teaching Hospital. A total of 544 data collected consisting of men between the age 17-86years with the mean aged group of 56years and was analysed using Epi-Info version 6.1. it was found that the prevalence of breast cancer among men was 4(2.6%), fibroadenoma197(36.8%),fibrocytic disease 120(22.4%), granulomatouse mastitis 14(2.6%)lactating adenoma 11(2.1%),sclerosing adenosis 8(1.5%), the highest prevalence rate was found between the aged group of 39-48years(50%) followed by 39-48years(25%) and 79-88years (25%) respectively.

KEYWORD: Breast Cancer, Male.

INTRODUCTION

Cancer is a term used for diseases in which abnormal cells proliferate without control.Breast cancer in men is a rare disease that accounts for less than 1% of all cancers in men and less than 1% of all diagnosed breast cancers,Magno et al,2009It is a diagnosis for which optimal management is not clearly established and treatment guidelines are scarce. The medical literature regarding breast cancer in men consists mainly of case-control and retrospective studies, and there are no randomised prospective data for this disease. Recent emphasis therefore has been placed on extrapolating data derived from studies of breast cancer in women and using those data as a benchmark for treating men—what’s good for the goose is good for the gander.Most types of cancer cells eventually form a lump; growth or mass called a tumor, and are named after the part of the body where the tumor originates. Breast cancer begins in breast tissue, which is made up of glands for milk production, called lobules, and the ducts that connect the lobules to the nipple. The remainder of the breast is made up of fatty, connective, and lymphatic tissue.Edge, et al,2010 Breast Cancer constitutes a major publichealth issue globallywith over 1 millionnewcases diagnosed annually, resulting in over 400,000annualdeath. Veronesi et al,2005, Omaret al,2013.There is an international/geographical variation in the incidence of Breast Cancer. Incidence rates are higher in the undeveloped countries than in the developing countries. Parkin ,et al,2005, Pages et al,2001. In Africa, Breast Cancer has overtaken cervical cancer as the commonest malignancy affecting women and the incidence rates appear to be rising. Vorobiofet al,2001, Omar et al,2013.In Nigeria breast cancer has increased from 116 per 100,000 in 2001. Adebamowo and, Ajayi. 2000. The activities of breast cancer society in some parts of the globe regarding cancer control and prevention may also be responsible for the better quality of live for victims of cancer in those countries Braunstein, 1993,Stratton et al,1997. Breast Cancers in developing countries are diagnosed when they are at advanced stages, which may be responsible for the higher mortality. The cost of treating Breast cancer in the tropics is prohibitive for most breast cancer victims; this is largely due to poverty and low per capita income as most people live below $1 per day, Thomaset al,1992, Althuiset al,1997.Cancers of cervix and breast have become preventable and have been well controlled In developed nations. Health education to increase awareness on breast self examination, may also contribute to early detection and prevention of invasive breast cancer,Adebamowo and, Ajayi ,2000. Agbo,et al,2013. The aim of this work is to determine the epidemiology and prevalence of breast cancer in menamong breast tissues submitted to histolopathology of university of Abuja Teaching Hospital, and also to determine the prevalence and proportion of breast cancer amongst the age group.

New Picture

This is a retrospective study comprises of 544 samples of breast tissue submitted to the histopathology laboratory of university of Abuja Teaching Hospital Gwagwalada Abuja Nigeria between 2010-2014.
Inclusion: Samples that have patient’s detail such as age, sex and diagnosiswere used
Exclusion: samples without age, sex ,and all females were excluded from the study.
Statistical analysis: Result of data were captured in excel and then analyzed using epi-info version 6.1

Methodology
Histological examinations
Breast tissues received registered, grossed and was processed in automatic tissue processor containing 12
beakers, The tissue was processed by allowing the tissue to passed through the following stages Fixation, Dehydration,Clearing,Impregnation,Embedding,Sectioning,Staining,Mountingthe tissues were passed from beaker 1,2,3 contains formalin where the tissues was allowed to stay for 30min each, beaker 4,5,6 contains alcohol and was allowed to stay for 1hour each, beaker 7,8,9 10 contains xylene and was allowed to stay for 1hr each while the last 2 beakers which is 11,and 12 contains wax and was allowed to stay in beaker 11 for 2hours (infiltration) and 12 for 2hours (impregnation).

Staining procedures for Haematoxylin and Eosin
Princples.
The haematoxylin is a basic dye and has affinity for the nucleus DNA & RNA which is acidic,the orange G6 has affinity for the acidophilic cells of the cytoplasm of superficial cells while the eosin azure has affinity for the cytoplasm of intermediate, parabasal and basal cells.

Methods
The smears was remove from fixatives and rinsed in descending grades of alcohol (80,70,50%),for 8secs each Stain in Harris alum-Haematoxylin for 4mins.
Wash in tap water for 1-2 mins, and was differentiated in 1% acid alcohol briefly.
It was then wash and blue in tap water for 3-5mins, and then transferred to two changes of 95% alcohol for a few seconds each.Stain in OG6 for 2minutes, Rinsed in two changes of 95% alcohol, and Stain in Eosin azure for 2-4mins.
It was rinsed in two changes of 95% alcohol .complete dehydration in absolute alcohol and cleared in xyleneand was mounted in DPX and examined to study the architecture of the tissues.
Any tissue examined and aberrations in the structural integrity of the tissues seen and a case of malignancy established, immunohistochemistry employed to profile the markers.

Principles of Immunohistochemistry
Immunohistochemistry (IHC) is a wide-used biological technique that combines anatomy, physiology, immunology and biochemistry. Developed from the antigen-antibody binding reaction, immunohistochemistry can be considered as a method that visualizes distribution and localization of specific antigen or cellular components in separated tissues, or tissue sections. Compared to other bio-techniques that are based on the antigen-antibody reaction such as immunoprecipitation, or western-blot, immunohistochemistry provides in situ information which promises a more convincing experimental result.
Major components in a complete immunohistochemistry experiment:
1) Primary antibody binds to specific antigen;
2) The antibody-antigen complex is formed by incubation with a secondary, enzyme-conjugated, antibody;
3) With presence of substrate and chromogen, the enzyme catalyzes to generate colored deposits at the sites of antibody-antigen binding.

New Picture (1)

Protocol

Prepare formalin-fixed, paraffin-embedded tissue sections (Step 1-8):
1. The freshly dissected tissue was fixed (<3mm thick) with 2% paraformaldehyde from 1h to overnight at room temperature.
2. The tissue was rinsed in running tap water for 5 min.
3. The tissue was then dehydrated through 70%, 80%, 95% alcohol, 5 min each, followed with 3 times of 100% alcohol, 5 min each.
4. The tissue was cleared in xylene for 2 times, 5 min each.
5. The tissue was immersed in paraffin for 3 times, 5 min each.

6. And Embeded in a paraffin block. The paraffin tissue block was then stored at room temperature for years.
7. The paraffin-embedded tissue block was section at 5-8 µm thickness on a microtome and floated in a 40°C water bath containing distilled water.
8. The sections were transferred onto glass slides suitable for immunohistochemistry. The slides were allowed to dry overnight and it was stored at room temperature until ready for use.
Immunostain formalin-fixed, paraffin-embedded tissue sections (Step 9-29):
9. The slides were Deparaffinizein xylene for 2 times, 5 min each.
10. The slides were transferred to 100% alcohol, for 2 times, 3 min each, and then transfer once through 95%, 70% and 50% alcohols respectively for 3 min each.
11. The endogenous peroxidase activity were blocked by incubating sections in 3% H2O2 solution in methanol at room temperature for 10 min to block endogenous peroxidase activity.
12. It was rinsed with PBS for 2 times, 5 min each.
13. The blocking buffer was drained from the slides.
14. 100 µl of diluted primary antibody was apply appropriately (in antibody dilution buffer, e.g. 0.5% bovine serum albumin in PBS) to the sections on the slides and incubate in a humidified chamber at room temperature for 1 h.
15. The slides were then washed with PBS for 2 times, 5 min each.
16. 100 µl of diluted biotinylated secondary antibody was apply appropriately (using the antibody dilution buffer) to the sections on the slides and incubate in a humidified chamber at room temperature for 30 min.
17. The slides were washed with PBS for 2 times, 5 min each.
18. 100 µl diluted Sav-HRP conjugates was apply appropriately (using the antibody dilution buffer) to the sections on the slides and incubate in a humidified chamber at room temperature for 30 min (keep protected from light).
19. The slides were washed with PBS for 2 times, 5 min each.
20. 100 µl DAB substrate solution was applied (freshly made just before use: 0.05% DAB – 0.015% H2O2 in PBS) to the sections on the slides to reveal the color of antibody staining.The color was allowed to development for < 5 min until the desired color intensity is reached. (Caution: DAB is a suspect carcinogen. Handle with care. Wear gloves, lab coat and eye protection.)
21. The slides were washed with PBS for 3 times, 2 min each.
22. The slides were Counterstain by immersing in Hematoxylin for 1-2 min.
23. The slides were rinsed in running tap water for 10 min.
24. Theslides were dehydrated through 4 times of alcohol (95%, 95%, 100% and 100%), 5 min each.
25. The slides were cleared in 3 times of xylene and coverslip using mounting solution. The mounted slides were stored at room temperature permanently.
26. The color of the antibody staining in the tissue sections were then observe under microscopy.

RESULT
Out of 544 data collected for breast tissues, 19 (3.6%) were males, Breast cancer showed a prevalence rate of 15

DISCUSSION
Of the 544 breast tissues submitted to histopathology laboratory of University of Abuja Teaching Hospital Gwagwalada, male study subjects shows a breast cancer prevalence rate of 4(2.6%) and the age range of the study subject 7-86,with the highest prevalence rate at 49–58yrs,(50%) followed by 39-48yrs and 79-88(25% and 25%) respectively which is agreement with a study carried out by (Kidman et al., 2000) in Jos with the age ranging from 12 -85 years which shows that Male breast cancer rate was 2% and also with 2.5% recorded in Benin by (Okobiaet al., 2001) and 1.47% recorded in Nnewi by (Anyanwu, 2000) respectively.
This result is not in agreement with the results obtained from Zaria (Hassan et al., 1995) and Jos which recorded 8.6% and 9% respectively. Also different from results obtained in Tanzania and Zambia which shows a prevalence rate of 6% and 15% respectively according to(Singgal et al, 2006) and (Ihekwaba, 1994). The high incidence of male breast cancer in Nigeria and Africa compare to Western countries have been attributed to hyperestrogenism due to endemic liver infections in Africa and environmental changes and lifestyle (Pere et al., 2007).
The definite etiology of male breast cancer is idiopathic just like other cancers. Factors such as alteration of hormonal milieus, family history and genetic alterations are known to affects its occurrence.Various studies have also shown that conditions that alter the estrogen-testosterone ratio in males predispose to breast cancer (Balleriniet al, 1990 and Casagrandeet al, 1988). Among these conditions, the strongest association is with Klinefelter Syndrome. Males with this condition have a fifty times increased risk and account for 3% of all breast cancer (Hultbornet al, 1997). Any condition associated with increased estrogen levels like cirrhosis and exogenous administration of estrogen (either in transsexuals or as therapy for prostate cancer) have been implicated as causative factors (Symmers, 1968, and Pritchard et al., 1988).
Androgen deficiency due to testicular disease like mumps, undescended testis, or testicular atrophy has been linked to the occurrence of breast cancer in men (Thomas et al., 1992) and (Mabuchi etal., 1985). Occupational exposure to heat and electromagnetic radiation, causing testicular damage and further leading to the development of male breast cancer have been postulated (Stenlund and Floderus, 1997) and (Pages et al., 2001). Hereditary breast cancers are known to occur in males.
Studies have found that gremlin mutations in BRCA1 and BRCA2 account most for this. (Stratonet al, 1997).Gynaecomastia has also been implicated as a risk factor (Braunstein, 1993).
The peak level incidence among Male was between 40 -86 years with mean age of 57 years which is similar to other results obtained in Nnewi, Nigeria(Anyanwu,2000) with a mean age of 60yrs and 66yrs obtained in Spain respectively (Ihekwaba,1994). KaiyumarContractor,et al,2008 also found average age mean of diagnosis to be 60yrs which is similar to the mean age of this study.
The burden of breast cancer among Nigerian women is becoming overwhelming, this may be due to the cultural practice in this country where most women seek alternative treatment and healing (ranging from herbal to religious help believing in the say that “It never my portion” or people attacking them in their villages and poverty rather than coming to die in the hospital. This partially explains why most women present with advance disease as our figure has shown (97.6%) thereby leaving them only the palliative option of treatment. Aggressive awareness campaign is the only way to change these attitudes.

CONCLUSION
The rise in breast cancer cases in this study population is an indication of inadequate or ineffective control measures to curtail the disease or due to diversion of global attention to HIV/AIDS and Tuberculosis in the country. Therefore, there is urgent need to step up activities through non-governmental agency to promote advocacy, national policy on training of personnel for clinical and self-breast examination, and nationwide screening program (Mammography) in order to enhance early detection, control the upward trends and reduce the mortality rate of breast cancer .Breast cancer aggressive awareness campaign should be increased to allow earlier detection so as to reduce the mortality and morbidity rate.

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