Detection of p53 Gene Status in Different Grades of Archival Breast Cancer Specimens by Quantitative Real-Time PCR
Author(s):
Maisaa Abdulmohsen Alwohhaib, Salah Khalid Al-Waheeb and Ghaneema Einad Alshammari
Background: Breast cancer is the most diagnosed cancer globally and contributes to the top five leading causes of cancer mortality. In Kuwait, breast cancer contributes for 49.4% from all types of cancers. P53 gene normal function is crucial in controlling cell proliferation and preventing the development of malignant cells. The aim of this study was to evaluate the p53 status in formalin fixed paraffin embedded (FFBE) samples of different grades of breast cancer using qPCR. The method of detecting p53 status affects the determination of its predictive value and the gene status present in the invasive forms of breast cancer. The aim of this study was to evaluate the p53 status in FFPE of different grades of breast cancer samples compared to the normal tissue by using qPCR. The method of detecting p53 status affects the determination of its predictive value and the gene status present in the invasive forms of breast cancer.
Methods: In this study 44 breast cancer tissue samples were used. DNA isolation was followed by quantitation of the HER2 gene which was done by real time PCR. The calculation of the relative fold values of the p53 gene was performed using the 2^ΔΔCt and ΔCt methods were the p53 gene was normalized with β actin gene. A one-way analysis of variance (ANOVA) test was used to determine whether the three groups of patients; grade I, II and III p53 qPCR relative values are statistically different.
Results: The ΔCt values for Grade I, Grade II and Grade III groups showed a noticeable down-regulation of the gene copy number values. However, the one-way ANOVA test P value for each of the 2^-ΔΔCt and ΔCt values were above the significance threshold 0.05. Accordingly, no significant differences among the three patient groups were found.
Conclusion: The down regulation of the gene observed in the invasive ducal and lobular carcinomas presented in the studied samples forms the start to investigate the p53 gene status in a larger group of patients. Determination of the p53 mutations is necessary and further studies are required to develop safe and efficient treatment to target p53 mutations in breast cancer.