Molecular and Immunohistochemical Study on Breast Cancer female Patients in a Sample of Iraqi Population

      Breast cancer is the most common cancer of women worldwide, Accounting for 25% of all cancers, with an estimated 1.57 million new cases in 2016. n Iraq,According to the latest Iraqi Cancer Registry, breast cancer is the most common malignancy among the Iraqi population THAT constitutes about one fourth of the registered cancer cases and it is the leading cause of death among the Iraqi population. The study aimed to detecting Immunohistochemical and molecular abnormalities in myeloid differentiation primary response gene 88 (MYD88) and Toll like Receptor 4 (TLR4) genes in Iraqi patients with breast carcinoma. The present study was conducted at AL- Emamain AL- Kadhemaian teaching Hospitals in Baghdad from April to November  2013 included 50 samples of female patient with malignant breast tumor, twenty samples of patient with benign breast tumor, and 25 blood samples from healthy individuals as controls. DNA extraction was performed from formalin fixed, paraffin embedded (FFPE) sections. After DNA quantification, the DNA samples were used for Conventional polymerase chain reaction (PCR) amplification of Toll like receptor 4 (TLR4) exon (1,2,3,4) by using five sets of primers  and Human myeloid differentiation factor 88 (MyD88) gene exon ( ,1 5) by using primer sets .the result revealed that The most of malignant BT patients were observed at the age group ≥ 40 years (66%), while in benign breast tumor patients were observed at the age group ≤ 40 years (60.0%).There is significant difference at malignant BT patients (P ≤ 0.01). agarose gel electrophoresis of the polymerase chain reaction (PCR) products of Toll like respecter 4 (TLR4) showed the presence of bands with size of 853 bp, 967 bp, 919 bp, 720 bp, polymerase chain reaction (PCR) products of myeloid differentiation primary response gene 88 gene showed the presence of bands with size of 606 bp, 877 bp.  PCR products were purified, and their DNA sequencing were performed, and analyzed by National Center for Biotechnology Information (NCBI). the result revealed that TLR4 have 21 mutations which are 12(57.14%) substitution, 7 (33.34%) deletion and 2 (9.52%) insertion. there is a significant difference among malignant BT patients groups (P<0.01). In myeloid differentiation primary response gene 88 gene the result revealed that substitution 3(100%). No insertion and no deletion in this gene. The differences in this study were significant (P<0.01) among malignant BT patients groups. Immunostaining was used to determine the positive expression of Toll like receptor 4 (TLR4). Among 30 samples of malignant breast tumor patient, 5(16.67%) have grad Ӏ, 13(43.33%) have grad ӀӀ, and 12(40%) have grad ӀӀӀ. difference (p<0.01) in these values in malignant BT patient. Immunohistochemical analysis of Toll like receptor 4 (TLR4) that conducted on 30 samples of malignant BT patient cleared that 24 (80%) patient had positive expression for Toll like receptor 4 (score +2&+3), while 6 out 30 samples (20%) scored +1, which is considered Toll like respecter 4 (TLR4) negative expression. Furthermore, 6 out of 15 (3.33%) samples from benign breast tumor scored +1, and 9 out of 15(60%) scored +2&+3.