الفهرس 
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Abstract
Breast cancer is the second most common diagnosed cancer overall but ranks 5th as cause of death. In Egypt, approximately 33% of all female cancer is breast cancer.
In the present study, histopathological, histochemical, and immunohistochemical studies as well DNA fragmentation assay were done on normal breast tissue, fibroadenoma and invasive ductal carcinoma tissues.
Patient groups
Sixty female breast specimens, obtained from Pathology Department, National Cancer Institute, Cairo Universtiy, were divided into three groups:
Group 1: Ten specimens were used as control; they were taken from adjacent fibroadenoma tissues.
Group 2: Ten specimens were diagnosed with breast fibroadenoma.
Group 3: Forty specimens were diagnosed with invasive ductal carcinoma.
Breast tissues from normal and breast cancer patients were fixed in 10% neutral formalin. After fixation, tissues were washed by running tap water over night. Specimens were dehydrated through ascending series of alcohol, then cleared in two changes of xylene and embedded in paraplast. Sections of 5 μm thickness were cut using rotary microtome for both histological, histochemical and immunohistochemical studies.
In histological studies, sections were stained in haematoxylin and eosin. For histochemical purposes, periodic acid Schiff (PAS) reaction was used for demonstration of polysaccharides, mercury bromophenol blue method for demonstration of total proteins and Schiff methylene blue for demonstration of DNA and RNA. For immunohistochemistry, antibodies directed against ER, PR, p53, Bax and Bcl-2. For DNA fragmentation tissues of normal, fibroadenoma and IDC were used.
The results can be summarized as follow
The mean age of female patients of control, fibroadenoma and IDC groups was 38.6 ± 3.931, 26.5 ± 8.38 and 46.17 ± 1.154 years, respectively. In IDC group, the number of patients with tumor size ˂ 2cm was 22 (55%) and number of patients with tumor size ≥2 cm was18 (45%). Number of patients with grade 1, grade 2, and grade 3 were none, 37, and 3, respectively.
Histopathological observations
Examination of normal breast sections stained with haematoxylin and eosin showed the lobule, the acini, the ducts and the stroma. Each acinus is enveloped by basement membrane and lined by two distinct types of cells arranged in spiral fashion; cuboidal or low columnar epithelial myoepithelial cells. The lobule was surrounded from outside and inside by connective tissue (stroma); interlobular and intralobular connective tissue, respectively.
Microscopic examination of fibroadenoma showed intracanalicular fibroadenoma, in which epithelial elements most commonly forms slit-like ductal structure and pericanalicular fibroadenoma, in which epithelial elements formed rounded or irregularly shaped tubules.
Examination of invasive ductal carcinoma showed that it consisted of irregularly shaped tightly or loosely cohesive groups, with pleomorphic tumor cells. The cells had moderate amounts of usually eosinophilic cytoplasm. The majority of the nuclei of these cells were irregular in shape and had different sizes.
Histochemical observations
Histochemical observations showed the presence of a noticeable difference in the content of polysaccharides, proteins and both types of nucleic acids (DNA
and RNA) in the three patient groups (normal, fibroadenoma and invasive ductual carcinoma). Normal breast tissues contained the highest polysaccharide contents, while fibroadenoma specimens showed moderate content and the lowest content was in IDC specimens.
Concerning the total proteins and nucleic acids, the invasive ductal carcinoma specimens showed significantly increase compared with fibroadenoma and normal specimens.
Immunohistochemical results:
There was highly significant increase in Bcl-2, Bax and p53 expression in invasive ductal carcinoma than other groups. The relationship between Bcl-2 and p53 expression was direct statistically significant, while the relationship between Bax and p53 expression was indirect statistically significant. There was a marked decrease in ER expression in both fibroadenoma and invasive ductal carcinoma than normal breast. The reduction of ER expression was highly significant at nvasive ductal carcinoma. In contrast, PR expression showed significantly increase in invasive ductal carcinoma group compared with normal breast group, while PR expression showed small decrease in fibroadenoma group than normal breast group. The relationship between p53 and ER or PR expression was indirect statistically significant.
DNA fragmentation results:
The total optical density in invasive ductal carcinoma specimens was significantly increased (98.2) compared with fibroadenoma specimens (0.3) and normal breast tissue specimens (0.6). These results indicated high DNA fragmentation in invasive ductal carcinoma.