Gynecomastia is often a known medical situation which causes males to have abnormal breast enlargement and growth - sometimes in just one breast and often in both. The actual condition tends to affect young boys going by means of puberty, and elderly men - because hormones often be out of balance in each stages of life. This is why so many want to findout how to get rid of gynecomastia.
When accurate Gynecomastia is happening to boys or males, there is about 1 as well as a half inches of location just beneath the nipple that tends to swell. It's often tender in this region also, and often even milk can be secreted.
There is no doubt that gynecomastia, or man breasts, is not only embarrassing and humiliating at occasions, but can even be painful also.This is why so many wat to know how to get rid of gynecomastia or how to lose man boobs. This medical condition affects males from puberty up to the elderly, and because of the obvious embarrassment, most people will not talk about it or seek out therapy for it. The truth is though, the problem is treatable.
How to Get Rid of Gynecomastia
The issue frequently goes away unaided on it is very own, within 1-3 years. Sometimes however, Gynecomastia is caused because of the man utilizing certain drugs which can cause the issue. It is a uncommon side impact of the drugs, but males can typically find how to get rid of Gynecomastia by basically changing the types of drugs they're taking, or utilizing alternatives.
Recreational drugs like marijuana and alcohol are recognized to cause Gynecomastia in uncommon cases for example, so you'll be able to get rid of the condition at times by basically stopping use of these. If you're taking certain prescription drugs on the other hand, including anything which creates additional estrogen, steroids, particular Cardiovascular drugs, and ulcer therapies for instance Tagamet, Prilosec, and Zantac - speak to your physician or medical care provider about making use of alternatives. You are able to generally get rid of Gynecomastia problems by basically stopping the use of those medicines, or altering to distinct sorts.
Now Gynecomastia is also referred to as "man boobs", and a lot more usually than not that particular dilemma is in fact caused by weight acquire and obesity. Getting rid of man breasts is becoming an obsession. This is just not accurate Gynecomastia though, instead this condition is known as Pseudogynecomastia: a.k.a. False Gynecomastia.
When a man has what appears to be "boobs" on his chest from straightforward weight acquire, it is relatively quick and uncomplicated to get rid of if you are prepared to place in a bit of work. You can't basically get rid of Gynecomastia or Pseudogynecomastia just by building up your chest muscles though. Actually, you can make the difficulty far more evident by building up the muscles.
Given that the fat and glands that are generating your boobs appear massive sit in between the muscles and your skin, building up the underlying muscles will merely push the "boobs" out further.
Discovering cures for gynecomastia is somewhat uncomplicated, but discovering 1 that truly works for you is the tough component. Take your time and appear at your selections in how to get rid of gynecomastia. By carrying out so you may uncover the right answer.
When the Gynecomastia is actually the false assortment caused by additional fat on the body, the most effective method to get rid of it is to start a fat burning routine. You will burn additional fat by toning up your muscles, but you'll want to select a diet and workout plan which is particularly created to assist burn excess fat from your body, then get started with it. Having a bit of dedication and commitment, you'll discover yourself simply in a position to get how to get rid of the Gynecomastia within just 3 to six weeks.
Discovering cures for gynecomastia is fairly straightforward, but discovering one that really functions for you will be the tough part. Take your time and appear at your possibilities in how to get rid of gynecomastia. By performing so you may discover the best remedy.
When accurate Gynecomastia is happening to boys or males, there is about 1 as well as a half inches of location just beneath the nipple that tends to swell. It's often tender in this region also, and often even milk can be secreted.
There is no doubt that gynecomastia, or man breasts, is not only embarrassing and humiliating at occasions, but can even be painful also.This is why so many wat to know how to get rid of gynecomastia or how to lose man boobs. This medical condition affects males from puberty up to the elderly, and because of the obvious embarrassment, most people will not talk about it or seek out therapy for it. The truth is though, the problem is treatable.
How to Get Rid of Gynecomastia
The issue frequently goes away unaided on it is very own, within 1-3 years. Sometimes however, Gynecomastia is caused because of the man utilizing certain drugs which can cause the issue. It is a uncommon side impact of the drugs, but males can typically find how to get rid of Gynecomastia by basically changing the types of drugs they're taking, or utilizing alternatives.
Recreational drugs like marijuana and alcohol are recognized to cause Gynecomastia in uncommon cases for example, so you'll be able to get rid of the condition at times by basically stopping use of these. If you're taking certain prescription drugs on the other hand, including anything which creates additional estrogen, steroids, particular Cardiovascular drugs, and ulcer therapies for instance Tagamet, Prilosec, and Zantac - speak to your physician or medical care provider about making use of alternatives. You are able to generally get rid of Gynecomastia problems by basically stopping the use of those medicines, or altering to distinct sorts.
Now Gynecomastia is also referred to as "man boobs", and a lot more usually than not that particular dilemma is in fact caused by weight acquire and obesity. Getting rid of man breasts is becoming an obsession. This is just not accurate Gynecomastia though, instead this condition is known as Pseudogynecomastia: a.k.a. False Gynecomastia.
When a man has what appears to be "boobs" on his chest from straightforward weight acquire, it is relatively quick and uncomplicated to get rid of if you are prepared to place in a bit of work. You can't basically get rid of Gynecomastia or Pseudogynecomastia just by building up your chest muscles though. Actually, you can make the difficulty far more evident by building up the muscles.
Given that the fat and glands that are generating your boobs appear massive sit in between the muscles and your skin, building up the underlying muscles will merely push the "boobs" out further.
- In some circumstances of gynecomastia, prescription drugs may be the trigger. In the event you have noticed adjustments inside your body since taking any new medicines, talk to your physician about it. They may be in a position to alter them up for you or make adjustments that may support.
- Your diet plan could be affecting the problem also. Are you overeating or overweight? By making modifications to your diet also can how to get rid of gynecomastia.
- Physical exercise that utilizes the chest muscles is yet another fantastic therapy in helping fight off the difficulty.
Discovering cures for gynecomastia is somewhat uncomplicated, but discovering 1 that truly works for you is the tough component. Take your time and appear at your selections in how to get rid of gynecomastia. By carrying out so you may uncover the right answer.
When the Gynecomastia is actually the false assortment caused by additional fat on the body, the most effective method to get rid of it is to start a fat burning routine. You will burn additional fat by toning up your muscles, but you'll want to select a diet and workout plan which is particularly created to assist burn excess fat from your body, then get started with it. Having a bit of dedication and commitment, you'll discover yourself simply in a position to get how to get rid of the Gynecomastia within just 3 to six weeks.
Discovering cures for gynecomastia is fairly straightforward, but discovering one that really functions for you will be the tough part. Take your time and appear at your possibilities in how to get rid of gynecomastia. By performing so you may discover the best remedy.
pre-sell article (part 2)
Introduction
Male breast cancer is an uncommon disease, accounting for approximately 1% of all breast cancer cases and less than 1% of all malignancies in men [1,2]. The American Cancer Society estimates that 1,910 men will be diagnosed with breast cancer in the USA in 2009 and about 440 men will die from the disease [3]. Although breast carcinoma in both genders shares certain characteristics, there are notable differences in incidence, age distribution, prognosis and survival. The unfavorable overall prognosis in male breast cancer has been attributed to the older age and advanced tumor stage at the time of diagnosis [4].
Based on the recent DNA microarray studies on female breast cancer cases, distinct molecular subtypes of breast carcinoma were identified with different clinical outcomes [5]. Using an intrinsic set of 534 genes, Sorlie and colleagues [6] analyzed the expression profiles of 115 independent breast cancer specimens and categorized them into the following subtypes: luminal; human epidermal growth factor receptor 2 (HER2) over-expressing; normal breast-like; and basal-like. The tumor subtypes correlated well with clinical outcomes as measured by overall survival and distal metastasis, with the worst outcome observed in HER2 over-expressing and basal-like subtypes [5-7].
The molecular subtypes of female breast cancer were originally identified by gene expression analysis using DNA microarrays. However, large-scale subtyping using gene expression profiling from formalin-fixed, paraffin-embedded samples is not currently feasible. Therefore, immunohistochemical (IHC) markers have been used as surrogates for DNA microarray in subtyping breast cancer. By using a panel of four antibodies including estrogen receptor (ER), HER1, HER2 and cytokeratin 5/6 (CK5/6), Nielsen and colleagues [8] characterized three IHC-defined subtypes: luminal (ER+, HER2-), HER2 (HER2+), and basal-like (ER-, HER2-, CK5/6+ or HER1+). Based on more recent gene expression studies, Carey and colleagues [9] updated IHC subtype definition as luminal A (ER+ and/or progesterone receptor (PR)+, HER2-), luminal B (ER+ and/or PR+, HER2+), HER2+/ER- (ER-, PR-, HER2+), basal-like (ER-, PR-, HER2-, CK5/6+), and unclassified (negative for all five markers).
In addition to these subtypes, the prognosis of breast cancer may be affected by expressions of two additional molecular markers. Epidermal growth factor receptor (EGFR) was reported to be over-expressed in aggressive female breast cancer [10-13] and is currently being evaluated in clinical trials as a potential therapeutic target [14-16]. It was also reported that transcription factor nuclear factor ?B (NF-?B) was activated in breast cancer cells with over-expressed EGFR [17]. The elevation of NF-?B was speculated to be associated with unfavorable prognosis by promoting metastasis [18,19], inhibiting apoptosis [20], and increasingly resistance to chemotherapy [21]. Increasing evidence has suggested that NF-?B is a potential target for the treatment of breast cancer and the prevention of metastasis [22,23].
Despite these advances in female breast carcinoma, our understanding and treatment strategies for male breast cancer are limited and generally extrapolated from our knowledge of female breast cancer. In particular, the molecular subtypes of male breast cancer have not been studied although the subtypes were reportedly associated with both biological and clinical behaviors in female breast cancer. This article reports our attempt to subclassify male breast carcinomas based on the immunoprofile, and to evaluate the association of the subtypes with histologic type, nuclear grade, lymph node status, clinical stage, and expression of EGFR and NF-?B.
Materials and methods
Specimens
Slides and paraffin-embedded tissue blocks from 42 male patients with breast cancer were retrieved from the surgical pathology archives in the departments of pathology of The University of Texas M. D. Anderson Cancer Center (from 2002 to 2005, Houston, Texas) and the University of Texas Medical Branch (from 1990 to 2004, Galveston, Texas). Clinical data for these patients were collected with approval of the Institutional Review Board (IRB# LAB05-0153).
Histologic evaluation
The slides from these cases were reviewed and the histologic diagnoses recorded in the medical record were confirmed independently by two pathologists (YMG, MG). The histologic classification was based on the criteria set by the World Health Organization. A diagnosis of invasive papillary carcinoma is rendered when a frankly invasive carcinoma predominantly in a papillary growth pattern, or in a ductal/solid patterns but associated with an in situ component of papillary carcinoma. The modified Black's nuclear grading system was used for nuclear grading with a three-tire scheme based on abnormalities in nuclear size, nuclear membrane, chromatin, and mitosis.
Immunohistochemistry and interpretation
Tissue sections (4 µm) from each case were prepared for immunostaining.
After incubation in a 60°C oven overnight and deparaffinization, the tissue sections were treated with 3% hydrogen peroxide in methanol for five minutes. Following a brief pretreatment with 0.02% protease XXIV for two minutes, the slides were incubated with one of the following antibodies: anti-human ER, PR, HER2, CK5/6, EGFR, and NF-?B. The sources and characterization of these antibodies are detailed in Table 1. The immunostains were performed using an automated staining system (BenchMark XT, Ventana Medical System Inc., Tucson, AZ, USA). Mouse Envision (DAKO Corp., St. Barbara, CA, USA) was used as secondary antibody. The color was visualized by incubation with chromagen DAB for 16 minutes. The slides were then counterstained with Mayer hematoxylin and a coverslip with Permount (Fisher Scientific, Fair Lawn, NJ, USA) was placed on the slide.
Table 1. Antibody characterization, dilutions and positive controls
The staining patterns and intensities for each of the markers were interpreted by two pathologists independently (YMG, MG). For ER and PR, nuclear staining in more than 10% of tumor cells was classified as positive staining. A positive HER2 stain was determined by greater than 2+ membranous staining of tumor cells based on the conventional three-tier grading criteria. All cases with greater than 2+ HER2 immunostain were further confirmed by fluorescence in situ hybridization (FISH) study. An estimation of more than 10% of tumor cells with membranous or cytoplasmic staining was required for a positive interpretation of EGFR and CK5/6. The NF-?B staining demonstrated a nuclear staining pattern, and if more than 10% of tumor cells stained, the specimen was considered to be positive.
FISH for HER2 amplification
The HER2 DNA probe kit (PathVysion; Vysis, Downers Grove, IL, USA) included two DNA probes directly labeled with different fluorescent dyes: the Spectrum-Orange fluorophore-labeled HER2 (190 kb) specific for the HER2 gene locus on chromosome 17q11.2-q12, and the Spectrum-Green fluorophore-labeled chromosome enumerator probe (5.4 kb) targeting the alpha satellite DNA sequence located at the centromeric region of chromosome 17 (CEP17; 17p11.1-q11.1).
FISH assay was performed according to the protocol provided by Vysis (Downers Grove, IL, USA). Routinely processed paraffin-embedded tissue sections (4 µm) were deparaffinized in three changes of fresh xylene for three minutes each, dehydrated in two changes of 100% ethanol for three minutes each, and then allowed to air dry. Slides were then placed in a preheated 80°C pretreatment reagent (1 M sodium isothiocyanate;Vysis, Downers Grove, IL, USA) for 13 minutes, rinsed in distilled water for three minutes, and allowed to air dry. Protease digestion was accomplished by placing the slides in a prewarmed 37°C protease solution (Vysis, Downers Grove, IL, USA) for 13 minutes. Samples were then rinsed in distilled water for three minutes and air-dried. All slides were hybridized under identical conditions and with appropriate control tissue. An aliquot of pre-diluted LSI HER-2 Spectrum-Orange/CEP17 Spectrum-Green (Vysis, Downers Grove, IL, USA) was applied to the region of interest on the slide. A coverslip was placed and sealed at the periphery with rubber cement. The slides were placed on a preprogrammed, humidified slide warmer (Hybrite; Vysis, Downers Grove, IL, USA) with the following settings: denaturation at 73°C for five minutes and hybridization at 37°C for 16 hours. After hybridization, the rubber cement was removed, and the coverslip was floated off by soaking the slides in two times standard saline citrate buffer with 0.3% Nonidet P-40 (2 × SSC/0.3% NP-40) at ambient temperature. The slides were incubated in pre-warmed 2 × SSC/0.3% NP-40 at 73°C for two minutes, immersed in 2× SSC/0.3% NP-40 at ambient temperature for one minute, air dried in the dark, counterstained with 7 ml of 4,6-diamidino-2-phenylindole dihydrochloride (DAPI; Vysis, Downers Grove, IL, USA), and a coverslip was placed on the slide.
The signals were enumerated using an epifluorescence microscope (Olympus AX70; Melville, NY, USA) fitted with a Spectrum-Orange, Spectrum-Green, and DAPI triple-filter set. At least 60 cells were scored in each preparation, and the copy numbers of HER2 and CEP17 for each cell were recorded. HER2 was quantified using the ratio of HER2 to CEP17 signal counts. HER2 gene amplification was defined as an HER2 to CEP17 signal ratio of 2.0 or greater. Polysomy of chromosome 17 was defined as the presence of three or more CEP17 signals in more than 6% of the tumor cells evaluated.
IHC subtyping criteria
The IHC-based definition of breast cancer subtypes used in this study was as follows: luminal A (ER+ and/or PR+, HER2-), luminal B (ER+ and/or PR+, HER2+), HER2+/ER- (ER-, PR-, HER2+), and basal-like (ER-, PR-, HER2-, CK5/6+) [9].
Statistical analysis
Descriptive statistics were calculated. Fisher's exact tests were used to assess the association between categorical variables. P values (two-sided test) of less than 0.05 were considered significant. All statistical analyses were carried out using SAS 8.0 (SAS Institute, Cary, NC, USA).
Results
The clinicopathological features of breast carcinoma from 42 men are summarized in Table 2. The racial distribution was as follows: 28 Caucasians, seven African Americans and seven Hispanics. Ages of the patients ranged from 33 to 82 years with a mean age of 63 years. Patients with luminal B subtype tumor was slightly younger than those with luminal A subtype (mean age 58 vs. 64 years, respectively), but the difference was not statistically significant (P = 0.25). Most of the patients were diagnosed with stage I to II disease (81%, 34/42) and were treated surgically (95%, 40/42), either by lumpectomy or mastectomy. Many patients also received other treatments including hormonal therapy (88%, 36/42), chemotherapy (64%, 27/42, with 2/27 received pre-operatively), and radiation therapy (19%, 8/42). There were no significant differences identified between the subtypes of male breast cancer regarding patients' race, presence of distant metastasis, and clinical stage.
Male breast cancer is an uncommon disease, accounting for approximately 1% of all breast cancer cases and less than 1% of all malignancies in men [1,2]. The American Cancer Society estimates that 1,910 men will be diagnosed with breast cancer in the USA in 2009 and about 440 men will die from the disease [3]. Although breast carcinoma in both genders shares certain characteristics, there are notable differences in incidence, age distribution, prognosis and survival. The unfavorable overall prognosis in male breast cancer has been attributed to the older age and advanced tumor stage at the time of diagnosis [4].
Based on the recent DNA microarray studies on female breast cancer cases, distinct molecular subtypes of breast carcinoma were identified with different clinical outcomes [5]. Using an intrinsic set of 534 genes, Sorlie and colleagues [6] analyzed the expression profiles of 115 independent breast cancer specimens and categorized them into the following subtypes: luminal; human epidermal growth factor receptor 2 (HER2) over-expressing; normal breast-like; and basal-like. The tumor subtypes correlated well with clinical outcomes as measured by overall survival and distal metastasis, with the worst outcome observed in HER2 over-expressing and basal-like subtypes [5-7].
The molecular subtypes of female breast cancer were originally identified by gene expression analysis using DNA microarrays. However, large-scale subtyping using gene expression profiling from formalin-fixed, paraffin-embedded samples is not currently feasible. Therefore, immunohistochemical (IHC) markers have been used as surrogates for DNA microarray in subtyping breast cancer. By using a panel of four antibodies including estrogen receptor (ER), HER1, HER2 and cytokeratin 5/6 (CK5/6), Nielsen and colleagues [8] characterized three IHC-defined subtypes: luminal (ER+, HER2-), HER2 (HER2+), and basal-like (ER-, HER2-, CK5/6+ or HER1+). Based on more recent gene expression studies, Carey and colleagues [9] updated IHC subtype definition as luminal A (ER+ and/or progesterone receptor (PR)+, HER2-), luminal B (ER+ and/or PR+, HER2+), HER2+/ER- (ER-, PR-, HER2+), basal-like (ER-, PR-, HER2-, CK5/6+), and unclassified (negative for all five markers).
In addition to these subtypes, the prognosis of breast cancer may be affected by expressions of two additional molecular markers. Epidermal growth factor receptor (EGFR) was reported to be over-expressed in aggressive female breast cancer [10-13] and is currently being evaluated in clinical trials as a potential therapeutic target [14-16]. It was also reported that transcription factor nuclear factor ?B (NF-?B) was activated in breast cancer cells with over-expressed EGFR [17]. The elevation of NF-?B was speculated to be associated with unfavorable prognosis by promoting metastasis [18,19], inhibiting apoptosis [20], and increasingly resistance to chemotherapy [21]. Increasing evidence has suggested that NF-?B is a potential target for the treatment of breast cancer and the prevention of metastasis [22,23].
Despite these advances in female breast carcinoma, our understanding and treatment strategies for male breast cancer are limited and generally extrapolated from our knowledge of female breast cancer. In particular, the molecular subtypes of male breast cancer have not been studied although the subtypes were reportedly associated with both biological and clinical behaviors in female breast cancer. This article reports our attempt to subclassify male breast carcinomas based on the immunoprofile, and to evaluate the association of the subtypes with histologic type, nuclear grade, lymph node status, clinical stage, and expression of EGFR and NF-?B.
Materials and methods
Specimens
Slides and paraffin-embedded tissue blocks from 42 male patients with breast cancer were retrieved from the surgical pathology archives in the departments of pathology of The University of Texas M. D. Anderson Cancer Center (from 2002 to 2005, Houston, Texas) and the University of Texas Medical Branch (from 1990 to 2004, Galveston, Texas). Clinical data for these patients were collected with approval of the Institutional Review Board (IRB# LAB05-0153).
Histologic evaluation
The slides from these cases were reviewed and the histologic diagnoses recorded in the medical record were confirmed independently by two pathologists (YMG, MG). The histologic classification was based on the criteria set by the World Health Organization. A diagnosis of invasive papillary carcinoma is rendered when a frankly invasive carcinoma predominantly in a papillary growth pattern, or in a ductal/solid patterns but associated with an in situ component of papillary carcinoma. The modified Black's nuclear grading system was used for nuclear grading with a three-tire scheme based on abnormalities in nuclear size, nuclear membrane, chromatin, and mitosis.
Immunohistochemistry and interpretation
Tissue sections (4 µm) from each case were prepared for immunostaining.
After incubation in a 60°C oven overnight and deparaffinization, the tissue sections were treated with 3% hydrogen peroxide in methanol for five minutes. Following a brief pretreatment with 0.02% protease XXIV for two minutes, the slides were incubated with one of the following antibodies: anti-human ER, PR, HER2, CK5/6, EGFR, and NF-?B. The sources and characterization of these antibodies are detailed in Table 1. The immunostains were performed using an automated staining system (BenchMark XT, Ventana Medical System Inc., Tucson, AZ, USA). Mouse Envision (DAKO Corp., St. Barbara, CA, USA) was used as secondary antibody. The color was visualized by incubation with chromagen DAB for 16 minutes. The slides were then counterstained with Mayer hematoxylin and a coverslip with Permount (Fisher Scientific, Fair Lawn, NJ, USA) was placed on the slide.
Table 1. Antibody characterization, dilutions and positive controls
The staining patterns and intensities for each of the markers were interpreted by two pathologists independently (YMG, MG). For ER and PR, nuclear staining in more than 10% of tumor cells was classified as positive staining. A positive HER2 stain was determined by greater than 2+ membranous staining of tumor cells based on the conventional three-tier grading criteria. All cases with greater than 2+ HER2 immunostain were further confirmed by fluorescence in situ hybridization (FISH) study. An estimation of more than 10% of tumor cells with membranous or cytoplasmic staining was required for a positive interpretation of EGFR and CK5/6. The NF-?B staining demonstrated a nuclear staining pattern, and if more than 10% of tumor cells stained, the specimen was considered to be positive.
FISH for HER2 amplification
The HER2 DNA probe kit (PathVysion; Vysis, Downers Grove, IL, USA) included two DNA probes directly labeled with different fluorescent dyes: the Spectrum-Orange fluorophore-labeled HER2 (190 kb) specific for the HER2 gene locus on chromosome 17q11.2-q12, and the Spectrum-Green fluorophore-labeled chromosome enumerator probe (5.4 kb) targeting the alpha satellite DNA sequence located at the centromeric region of chromosome 17 (CEP17; 17p11.1-q11.1).
FISH assay was performed according to the protocol provided by Vysis (Downers Grove, IL, USA). Routinely processed paraffin-embedded tissue sections (4 µm) were deparaffinized in three changes of fresh xylene for three minutes each, dehydrated in two changes of 100% ethanol for three minutes each, and then allowed to air dry. Slides were then placed in a preheated 80°C pretreatment reagent (1 M sodium isothiocyanate;Vysis, Downers Grove, IL, USA) for 13 minutes, rinsed in distilled water for three minutes, and allowed to air dry. Protease digestion was accomplished by placing the slides in a prewarmed 37°C protease solution (Vysis, Downers Grove, IL, USA) for 13 minutes. Samples were then rinsed in distilled water for three minutes and air-dried. All slides were hybridized under identical conditions and with appropriate control tissue. An aliquot of pre-diluted LSI HER-2 Spectrum-Orange/CEP17 Spectrum-Green (Vysis, Downers Grove, IL, USA) was applied to the region of interest on the slide. A coverslip was placed and sealed at the periphery with rubber cement. The slides were placed on a preprogrammed, humidified slide warmer (Hybrite; Vysis, Downers Grove, IL, USA) with the following settings: denaturation at 73°C for five minutes and hybridization at 37°C for 16 hours. After hybridization, the rubber cement was removed, and the coverslip was floated off by soaking the slides in two times standard saline citrate buffer with 0.3% Nonidet P-40 (2 × SSC/0.3% NP-40) at ambient temperature. The slides were incubated in pre-warmed 2 × SSC/0.3% NP-40 at 73°C for two minutes, immersed in 2× SSC/0.3% NP-40 at ambient temperature for one minute, air dried in the dark, counterstained with 7 ml of 4,6-diamidino-2-phenylindole dihydrochloride (DAPI; Vysis, Downers Grove, IL, USA), and a coverslip was placed on the slide.
The signals were enumerated using an epifluorescence microscope (Olympus AX70; Melville, NY, USA) fitted with a Spectrum-Orange, Spectrum-Green, and DAPI triple-filter set. At least 60 cells were scored in each preparation, and the copy numbers of HER2 and CEP17 for each cell were recorded. HER2 was quantified using the ratio of HER2 to CEP17 signal counts. HER2 gene amplification was defined as an HER2 to CEP17 signal ratio of 2.0 or greater. Polysomy of chromosome 17 was defined as the presence of three or more CEP17 signals in more than 6% of the tumor cells evaluated.
IHC subtyping criteria
The IHC-based definition of breast cancer subtypes used in this study was as follows: luminal A (ER+ and/or PR+, HER2-), luminal B (ER+ and/or PR+, HER2+), HER2+/ER- (ER-, PR-, HER2+), and basal-like (ER-, PR-, HER2-, CK5/6+) [9].
Statistical analysis
Descriptive statistics were calculated. Fisher's exact tests were used to assess the association between categorical variables. P values (two-sided test) of less than 0.05 were considered significant. All statistical analyses were carried out using SAS 8.0 (SAS Institute, Cary, NC, USA).
Results
The clinicopathological features of breast carcinoma from 42 men are summarized in Table 2. The racial distribution was as follows: 28 Caucasians, seven African Americans and seven Hispanics. Ages of the patients ranged from 33 to 82 years with a mean age of 63 years. Patients with luminal B subtype tumor was slightly younger than those with luminal A subtype (mean age 58 vs. 64 years, respectively), but the difference was not statistically significant (P = 0.25). Most of the patients were diagnosed with stage I to II disease (81%, 34/42) and were treated surgically (95%, 40/42), either by lumpectomy or mastectomy. Many patients also received other treatments including hormonal therapy (88%, 36/42), chemotherapy (64%, 27/42, with 2/27 received pre-operatively), and radiation therapy (19%, 8/42). There were no significant differences identified between the subtypes of male breast cancer regarding patients' race, presence of distant metastasis, and clinical stage.
