ظافر عبد المهدي الكوفي ( أستاذ مساعد )
كلية الصيدلة - صيدلة عام
[email protected]
A Biochemical Study of FOXM1 and TP53 Genes in Breast Cancer Patients by RT-qPCR
بحث النوع:
طب التخصص العام:
Dhafer A.F. Al-Koofee اسم الناشر:
اسماء المساعدين:
Lambert الجهة الناشرة:
2015 سنة النشر:


Background: Breast cancer is a malignant tumor arising from the terminal duct lobular unit of the breast tissues. It is most prevalent malignancy in women and its incidence is increasing, approximately equals to 5-10%. Forkhead box M1 (FOXM1) is a transcription factor associates with proliferation signals and over-expressed in majority of human cancer involved breast. TP53 gene negatively regulates FOXM1 expression and therefore inactivation of TP53 in tumors could partially illustrate the phenomenon of FOXM1 over-expression in human cancers. Objectives: The main objective of the current study is to analyze FOXM1 and TP53 gene expressions levels in formalin fixed paraffin embedded tissues of breast cancer specimens. Subjects and Methods: One hundred formalin fixed paraffin embedded (FFPE) tissue samples of malignant breast tumors as well as 100 FFPE samples of normal breast tissues were investigated. Ages of patients (Mean ± SD) were 48.4 ± 10.90 year with a minimum to maximum of 20 -75 years. Total mRNA was extracted from FFPE blocks by using a specific kit. FOXM1 and TP53 gene expressions were estimated quantitatively by real-time PCR. Results were normalized to 18S rRNA gene as housekeeping gene. The gene expression data were analyzed in relevance to the patients information obtained. Several statistical analyses were applied to analyze the data. Results: Analysis of the data of the ages revealed 29 (29%) patients of ≤40 years and 71 (71%) of >40 years. Ninety one cases (91%) out of 100 were found to be invasive ductal carcinomas, 7 cases (7%) out of 100 were invasive lobular carcinoma, and 2 cases (2%) out of 100 were rare type of breast cancer. Tumor size assessment indicated 10 (10%), 51 (51%), 26 (26%), and 13 (13%) cases to be of T1, T2, T3, and T4 respectively. Lymph node involvement was positive in 77 (77%) and negative in 23 (23%) cases. Fifty three (53%) cases were of right sided and 47 (47%) cases were of left sided. Estrogen receptors were found to be positive in 44 cases (44%), negative in 53 cases (53%) and unknown in 3 cases. Progesterone receptors were observed to be positive in 50 cases (50%), negative in 47 cases (47%) and undefined in 3 cases (3%). Her2 was exhibited to be positive in 16 (16%) cases, negative in 76 (76%) cases, and unknown in 8 cases (8%). The concentration (Mean ± S.D) of extracted total RNA from FFPE samples was found to be 55.5±13.61 µg/ml with a purity of 1.93±0.04. Extracted RNAs were converted to cDNA and amplified with the use of two levels (5 and 10 pmole) of the designed primers. Both levels were observed to amplify the cDNA successfully. The specificity of the designed primer was confirmed by monitoring the melting curve of the amplified DNA. FOXM1 and TP53 cDNAs were amplified successfully with 99% and 78% PCR efficiency respectively. The expression folds of FOXM1 and TP53 genes were found to be 15 and 44 folds in malignant breast tumors respectively. High expression folds of FOXM1 and TP53 genes were evident in tumor of patients with ages ≤40 and >40 years respectively. FOXM1 and TP53 genes were observed to be expressed excessively in ductal carcinoma tumors, advanced stage tumors, estrogen and progesterone receptors negative tumors, and those positive for Her2.