الفهرس 
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Abstract
Cell proliferation can be stimulated by physiologic and pathologic conditions requiring the selective expression of biological markers involves a complex and interdependent series ofnuclear structureand specific genesfrom the microenvironment which either stimulate or inhibit.
The current study was undertaken on 40 prospective biopsies taken from patients with breast tumor. They were divided into benign (10 cases) and malignant (30 cases) groups as well as control (10 cases taken from tissue parenchyma adjacent to the tumor). All biopsies were obtained from Department of Pathology, Medical Research Institute, and Alexandria University. During the period from December 2014 to Mares 2015.
Gross examination for surgically excised samples was done with the measurement of the tumor size and axillary lymph nodes dissection. Parts from the tumors and from the tissue adjacent to the tumor were subjected to the standard procedure of fixation, dehydration, paraffin embedding and sectioning and then subjected to Haematoxlylin and Eosin (H&E) stain for histological and histopathological structure demonstration, Feulgen staining for DNA demonstration, Ag NORs silver-staining for demonstrating the argyrophilicnucleolar organizer region andimmunohistochemical staining for ki-67 protein as aproliferatingmarker
The present study aimedto assess the proliferation induce through histochemical and immunohistochemical studies and evaluate the correlation ofproliferationmarkersinboth benign and malignant breastlesions with hormonal receptor status and other clinicopathological parameters.
The current results represent a percentage histopathological evaluation of the breast lesion. The benign groups represent fibroadenomas (60%, 6/10) and (40%, 4/10) fibrocystic diseases.The malignant groups represent (90%, 27/30) of invasive ductal carcinoma (IDC) and (10%, 3/30) invasive lobular carcinoma (ILC). The invasive ductal carcinoma were diagnosed as (66.7%, 18/27) grade II and (33.3%, 9/27) grade III.
The other pathological parameter represent a majority tumor sit affected is the left tumor mass(LT), the majority of tumor size among2≤5cm in the malignant cases and the age was distributed at15-25 yearsin benign casesand55-65 yearsin malignant cases. Also, the positive lymphatic metastasis was distributed in 90% of all malignant cases.
In addition, estrogen receptor (ER) immunostainingwas distributed a markedpositive in44.4% of grade II and, (50.0%)ofgrade III, while the progesterone receptor( PR) immunostaining was distributedweak positive in 38.9% of grade II and 50% of grade III. Also, the immunostaing of Her2 positive was distributed as a strong positive in grade II as 33.3%and25%ofgrade III.
The present study dependent on the histochemical and immune-histochemical assessment for detected the nuclear protein which related to DNA replication.The histochemical detection was used:
1. Feulgen stain for assessing DNA content in the ductal breastcells nuclei. The results evaluated the deep magentacolor in normal individual case and pale magenta color in malignant due the decreased DNA content..
2. The silver stain for assessing the nuclear organizer region Ag(NORs) in the ductal breast cells. The results revealed the increased of the dark and size black dots in nucleoli and nuclear matrix in the malignant group.There was a statistical significant increase (P ≤ 0.001*) in the histochemical of Ag NOR staining in malignant and benign individual than in control.
3. In addition, the immunohistochemical detection of Ki67 protein which is produced in the proliferative phase of the cell cycle. The results appeared the high expression of the Ki67 positive nuclei countsat the anaphase compare to the metaphasewith significant correlation (p ≤ 0.001) in both benign and malignant cases. Also, the positive nuclear count of the Ki67 expression was noticed in the grade III of the tumor cases with a significant difference (p ≤ 0.001) correlated to thegrade II.
The new concept of this work was quantitatively the assessment of the histochemistry and immunohistochemistry stains caries for calculating density of stains by image optical density (IOD) using digital image analysis software. The mean values of each reaction were based on the mean of pixel digit. The IOD intensity color based on Gray value -level differentiatedin digitized imagesaccording to different reactionfrom dark to light(140down in 70 ) in Feulgen.( 132.18 down82.94) in Ag(NORs)(104.67up to 137.54) in Ki67 (75.69 up to 140.04).
The present results of the image analysis (IOD) revealed that:
• The possibility of quantifying DNA by the Feulgen stain is proportional to the DNA concentration associated to the histone protein( chromosome), which present a high mean evaluation in the control than to the tumor individuals with a significant decreased (P ≤ 0.001*). Aw well as was noticed at the high grade of tumor metastasis.
• The proliferation induce assessmentby AgNOR and Ki67 antibody represent that the IOD evaluation elevated a high significant (P≤ 0.001*) increased in proliferative activity positive stain density (IOD) in both AgNOR and Ki67 in benign and malignant cases as well as the high grade tumor. Whereas, there was no significant difference between the both result of quantitative studies( count and IOD)in breast lesion cases.The two markers were calculated in the area of nucleoli and nucleus that thestains wereestimated in nuclear proliferating area not in the cytoplasm.