title: Resistance to spontaneous apoptosis in acute myeloid leukaemia blasts is associated with p-glycoprotein expression and function, but not with the presence of FLT3 internal tandem duplications.
abstract: The ability of acute myeloid leukaemia (AML) blasts to survive in culture has been associated with poor patient response to chemotherapy. Other biological factors predicting an adverse outcome include p-glycoprotein (pgp) expression, which is associated with a reduced remission rate, and the presence of fms-like tyrosine kinase 3 gene (FLT3) internal tandem duplications (ITDs), predictive of a high rate of leukaemic relapse. Our previous work has indicated a drug efflux-independent role for pgp in apoptosis resistance. We measured spontaneous in vitro apoptosis in 58 primary AML samples to establish its relationship with functional and phenotypic pgp and with FLT3 ITDs. Cells were incubated for 48 h in a suspension culture, and the remaining viable cells were counted by flow cytometry. Median survival was 38% of baseline values. Resistance to spontaneous apoptosis was strongly associated with pgp (MRK-16 antibody) expression (P = 0.001) and with pgp functional activity (P < 0.001). FLT3 ITDs, found in 20 cases, were inversely associated with functional pgp activity: thus, the median pgp modulation ratio was 2.0 in FLT3 wild-type cases and 1.38 in ITD cases (P = 0.018). Also, the presence of FLT3 ITDs was not associated with in vitro apoptosis resistance. In conclusion, we have found that the presence of FLT3 ITDs is not related to AML blast survival in vitro, and is inversely associated with pgp activity, whereas pgp expression and activity are associated with resistance to spontaneous apoptosis. These results may help to explain the differing adverse effects of pgp (on remission induction) and FLT3 ITDs (on relapse) in AML.
authors.affiliation: Division of Haematology, University of Nottingham and Nottingham City Hospital, Nottingham, UK. monica.pallis@nottingham.ac.uk
authors.forename: Monica
authors.initials: M
authors.lastname: Pallis
authors.forename: Julie
authors.initials: J
authors.lastname: Turzanski
authors.forename: Martin
authors.initials: M
authors.lastname: Grundy
authors.forename: Claire
authors.initials: C
authors.lastname: Seedhouse
authors.forename: Nigel
authors.initials: N
authors.lastname: Russell
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000208:Acute Disease; D017209:Apoptosis; D003524:Cyclosporins / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D007951:Leukemia, Myeloid / Q000378:metabolism / Q000473:pathology*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D012008:Recurrence; D012074:Remission Induction; D012091:Repetitive Sequences, Nucleic Acid; D020112:Rhodamine 123; D018709:Statistics, Nonparametric; D014407:Tumor Cells, Cultured; D051941:fms-Like Tyrosine Kinase 3
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D003524:Cyclosporins; D005456:Fluorescent Dyes; D011518:Proto-Oncogene Proteins; D020112:Rhodamine 123; C497970:FLT3 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3; C070272:valspodar