EVALUATION OF DNA ANALYSIS FOR EVIDENCE OF APOPTOSIS IN MEGALOBLASTIC-ANEMIA

This study involved DNA analysis of bone marrow cells of 15 patients w ith megaloblastic anaemia. The diagnosis was based on the morphologica l changes seen in the bone marrow, associated with either a low red ce ll folate or serum vitamin B12 level and an adequate response to appro priate therapy as confirmation of the diagnosis. Flow cytometric DNA a nalysis showed an increase in the S and G(2) phases of the cell cycle, but conventional agarose gel electrophoretic DNA analysis did not con firm the characteristic 'ladder pattern' which might have been expecte d in classic apoptosis. In addition, cells showing morphological chang es suggestive of apoptosis, such as nuclear condensation and fragmenta tion, did not show evidence of DNA fragmentation using the ApopTag(TM) in situ digoxigenin nucleotide labelled, peroxidase detection system, Further studies using annexin V now cytometric analysis and pulsed fi eld gel electrophoresis were also unable to detect evidence of apoptos is as a significant cause of cell death in megaloblastic anaemia.