The study of mutations of CDK4 and TP53 genes in selected breast lesions

Abstract

Lesions are abnormal changes in tissue. The theories behind tumorigenesis can be genetic epigenetic, immune surveillance and monoclonal hypothesis, which led to genetic changes. The genetic change can be the inactivation of tumor suppressor genes or the activation of oncogenes in the cell cycle. This study investigated the presence of gene mutation in CDK4 and TP53 genes in breast lesions. Ten (10) formalin fixed paraffin embedded tissue block, which includes 5 cases of fibroadenoma and 5 cases of invasive ductal carcinoma from the pathological archives were used. The nuclei amplification technique was carried out on the formalin fixed paraffin embedded tissue blocks. DNA extraction involves lysing the cells and solubilizing DNA, which is followed by chemical or enzymatic methods to remove macromolecules, lipids, RNA or proteins. Thereafter, the DNA is amplified using reagents and heated (denaturing) and cooling (annealing) steps. PCR requires free nucleotide (dNTPs), template DNA to amplify and unique single-stranded DNA primers that bind upstream (5') and downstream (3') of the DNA region of the tissue blocks. The integrity of the DNA is assessed by loading approximately 100 ng per sample on a 1.5% agarose gel and the sizes of PCR products were estimated by comparison with the mobility of a 100bp molecular weight ladder that was run alongside the samples in the gel. However, the amplified fragment was purified to remove PCR reagents and thereafter it was sequenced. The sequence of single nucleotide polymorphism of each gene was analyzed, and the results were reported. In mutation was more in both genes compared to the sequence of FA and IDC. From this study, both genes showed segments of mutation which were either SNPs or functional mutation. It showed that both genes were modified from a proto-oncogene to an oncogene and the protein it produces is an oncoprotein and as such it favours the alteration of both genes led to cell proliferation. The data generated from this research has highlighted the importance of carrying out further studies in assessing both genes to understand their involvement in the etiology of carcinogenesis which will help in clinical information about breast cancer. In line with the findings, there is an urgent need to understand carcinogenesis which may serve in early diagnosis and understanding of breast conditions and further help prevent or delay the onset of chronic complications and reduce morbidity and mortality rates to improve and assure the health of individuals.