Author name:
هبة لفتة عبد الغالبي
Supervisor name:
منال بادي صالح | مها شاكر حسن
Abstract:
Breast cancer is a complex and multifactorial disease resulting in abnormal cell growth that leads to malignant tumor formation.In this study we focus on mutations of breast cancer susceptibility genes 1 and 2 (BRCA1 and BRCA2), immunohistochemical evaluation of estrogen, progesterone, HER - 2 receptors, ABO blood groups , white blood cells and clinicopathological parameter ( age, breast feeding, family history and tumor site) .Eighty five blood samples were taken from patients who attended Al - Hussain teaching hospital and oncology unit in Al - Habboby hospital during the period (from August /2014 to April /2015) , fifty blood samples were collected from healthy women as a control group who attended Al - Hussain teaching hospital and divided into groups according to age, family history, age at first brith, breast feeding. For histopathological and immunohistochemical analysis fifty tissue samples were collected from patients with breast cancer who were undergoing surgical resection (mastectomy). Blood samples were obtained by venepuncture , using a 5 ml disposable syringe , and they were divided into three aliquots, which were 1 ml, 2 ml and 2ml, for collect serum, genetic studies and hematological parameters (leucocycte count and ABO blood groups) study. The tissue samples were collected from the 50 patients with breast cancer for histopathological diagnosis and for Immunohistochemical analysis (estrogen receptor, progesterone receptor and here2/nue) The total patients were divided according to their age in to five groups (30 - 39), (40 - 49), (50 - 59), (60 - 69) and (≥70 years). results of present study revealed that a highest incidence of breast cancer (41%) occur in the age oup between 40 - 49 years, there is a significant difference among patients groups (P ≤ 0.01).The results of our study showed that (77) of patients lactating and from those according to period of lactation we found that the highest percentage of patients (37.6%) were lactating for period between (18 - 22months) and the lowest percentage (10.38%) lactating for period between (1 - 6 month). Family history of breast cancer indicates a strong association with risk of developing breast cancer. The result of the present study revealed that the family history positive in 31 cases (36.47%), and 54 cases (63.52%) had negative family history of breast cancer.The results of present study showed that (41.5%) had first birth at age (20 - 29years), (23.37%) had their first birth in age group (15 - 19 years) and (30 - 39 years), and (9.09%) had first birth at age (≥40 years).Regarding the tumor site, we found that most of patients (50.5%) the tumor mass was located in the right side and (38.8%) cases the tumor mass were located in the left side.The DNA was extracted from blood samples by using Accupower® genomic DNA extraction kit. Each concentration and purity of all DNA samples have been measured by Nanodrop. Samples with a purity ranged from 1.2 to 1.8 have been enrolled in this study for the molecular detection of BRCA1/2 gene mutation in patients and control groups. Detection of BRCA1/2 gene mutation by multiplex PCR revealed that (185 del AG mutation in BRCA1 gene) was detected in 6 patients (7.05%) out of 85 cases with breast cancer, and (5382 ins C) mutation in the same gene was detected in 2 patients (2.35%). Regarding (6174 del T) mutation in BRCA2 gene was detected in 3 patients (3.52%). While this mutation was not detected in the control groups.Histopathologically, carcinoma was divided into 44 cases (88%) ductal carcinoma {from which 42 cases (84%) were invasive ductal carcinoma, and 2 cases (4%) were comedocarcinoma}, and 6 cases (12%) cases invasive lobular carcinoma. Results revealed that 2 cases (4%) were stage I, 13 cases (26%) were stage II, 24 cases (48%) were stage III and 11 cases (22%) were stage IV, also our results showed that the 2 cases (4%) were grade I, 23 cases (46%) were grade II and 25 cases (50%) were grade III. The results of immunohistochemical analysis for breast cancer samples showed that Estrogen receptors were present in 36 cases (72%), and 14 cases (28%) were negative, while Progesterone receptors detected in 27 cases (54%), and 23 cases (46%) were negative, on the other hand the expression of HER - 2/neu showed positive results in 28 cases (56%), and 22 cases (44%) were negative.Regarding the ABO blood groups, we found that most of the cases carry blood group A (40 cases, 47.05 %), and the lowest frequency was for blood group AB (7 cases, 8.23%).The results of the present study showed that leukocytes count, lymphocytes count, monocytes count, neutrophils count and basophiles count were lower in patients than the control group, with a significant difference (p value ≤ o.o1). While the eosinophils count was significantly higher in patients than the controls (p value ≤ o.o1).
