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Evaluation of DNA analysis for evidence of apoptosis in megaloblastic anaemia
Author(s) -
Ingram C. F.,
Davidoff A. N.,
Marais E.,
Sherman G. G.,
Mendelow B. V.
Publication year - 1997
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1997.d01-2075.x
Subject(s) - dna fragmentation , agarose gel electrophoresis , apoptosis , bone marrow , microbiology and biotechnology , biology , fragmentation (computing) , dna , programmed cell death , pathology , megaloblastic anemia , vitamin b12 , immunology , biochemistry , medicine , ecology
This study involved DNA analysis of bone marrow cells of 15 patients with megaloblastic anaemia. The diagnosis was based on the morphological changes seen in the bone marrow, associated with either a low red cell folate or serum vitamin B12 level and an adequate response to appropriate therapy as confirmation of the diagnosis. Flow cytometric DNA analysis showed an increase in the S and G 2 phases of the cell cycle, but conventional agarose gel electrophoretic DNA analysis did not confirm the characteristic ‘ladder pattern’ which might have been expected in classic apoptosis. In addition, cells showing morphological changes suggestive of apoptosis, such as nuclear condensation and fragmentation, did not show evidence of DNA fragmentation using the ApopTag TM in situ digoxigenin nucleotide labelled, peroxidase detection system. Further studies using annexin V flow cytometric analysis and pulsed field gel electrophoresis were also unable to detect evidence of apoptosis as a significant cause of cell death in megaloblastic anaemia.