Detection of p53 protein in leukemic cells by flow cytometry and Western blot

Abstract

Introduction: The p53 protein plays a crucial role in the cell cycle control, DNA damage repair and induction of apoptosis in genetically unstable cells. The Western blot is the most utilized methodology for p53 detection, but it requires lots of time to be executed and is a very elaborated technique. Nowadays, flow cytometry (FC), a more practical technique is being used for p53 detection. Objectives and methodology: to compare the results of p53 detection by FC and WB in leukemic cells. We used samples from 3 patients with acute lymphoid leukemia (ALL), 5 with acute myeloid leukemia (AML), 6 with chronic myeloid leukemia (CML) and 8 with chronic lymphoid leukemia (CLL). The positive (5) and negative (4) controls, used in both methods were leukemic cells lineages. Lymphocytes from 40 healthy donors were used as control for negative labeling in FC. The FC was performed after labeling with p53 monoclonal antibody and the WB by usual protocol. Results and conclusion: We verified agreement in the results in 82% of leukemic samples and in 100% of cell lineages. FC+/WB+ results were observed in patients with poor prognosis such as in CLL / Richter Syndrome, CML in blastic crisis and in the majority of AML. Although WB is considered as standard methodology for p53 detection, our results show that FC can be satisfactorily used for detection of this protein in leukemic cells.