Expression Analysis of Caspase-3 (CASP3) Gene in Leukemia Patients Using Quantitative Polymerase Chain Reaction (qPCR) and Western Blot Techniques

Abstract

CASP3 belongs to the caspases family and accounts for several critical activities, including apoptosis and other pathological and physiological processes. CASP3 also influences the multiple functions in cellular development such as homeostasis, hematopoietic growth and embryonic growth. It has significant involvement in the initiation and development of cancers including breast, colon and uterine cancers. However, molecular studies on the CASP3 gene in leukemia have not been widely conducted. The present study was designed to analyse the expression level of the CASP3 gene in leukemia patients. The results were further compared between different parameters such as types of leukemia, age, gender, occupation, WBC count and location-based. About 112 leukemia blood samples along adjacent controls were collected to analyse the CASP3 expression in this study cohort. The mRNA of each patient and control samples were isolated, quantified and converted to complementary DNA (cDNA). Relative expression of the CASP3 gene was measured using the quantitative PCR (qPCR) and Western blot analyses. Results showed that the relative expression of CASP3 was found to be significantly downregulated at both mRNA and protein levels in leukemia patients compared to normal controls. Further analysis showed that expression of CASP3 was downregulated in patients receiving the combination of chemotherapy as well as radiotherapy compared with patients receiving chemotherapy. These findings from our research showed the lower expression of CASP3 is associated with an increased risk of development and proliferation of leukemia. It can, therefore, be concluded that lower expression of the CASP3 gene may contribute to the initiation and development of leukemia.