Cellular signaling events elicited by v-abl associated with growth factor independence in an interleukin-3-dependent cell line

Owen, P J; Musk, Philip; Evans, Caroline A.; Whetton, Anthony D.

doi:10.1016/s0021-9258(18)82312-2

Cited by 29 publications

(4 citation statements)

References 43 publications

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“…2-Deoxy-n-Glucose (2-DOG) Uptake Assay. Cells were taken from mid log-phase cultures grown for 18 h at 39~ to ensure inactivation of v-ABL (21,22). They were then washed three times over a period of 2 h at 39~ in Fischer's medium containing 5 mM Hepes, pH 7.4, to remove IL-3.…”

Section: Methodsmentioning

confidence: 99%

“…Cell Culture and Reagents. IC.DP cells were maintained in Fischer's medium supplemented with 10% (vol/vol) horse serum and 3% (vol/vol) Ib3-containing X63-Ag-653 cell-conditioned medium as previously described (21,22).…”

Section: Methodsmentioning

confidence: 99%

“…Detection of Phosphorylation of the p160 ~*bl Tyrosine Kinase. 321)_ labeled cell lysates were immunoprecipitated with polyclonal antibodies against c-ABL (Cambridge Bioscience, Cambridge, UK), followed by SDS/polyacrylamide gel electrophoresis and autoradiography using previously described methods (21,22).…”

Section: Methodsmentioning

confidence: 99%

“…In view of the abrogation of the survival requirement for IL-3 by activation of v-ABL, we assessed the effects of v-ABL activity on glucose uptake to determine the relationship, if any, between glucose transport and apoptotic suppression. We have previously shown that activation of v-ABL suppresses apoptosis and thus prolongs the survival, but does not allow the proliferation, of IC.DP cells in the absence of growth factor (21,22). Here we demonstrate that v-ABL activation suppresses the onset of apoptosis in IC.DP cells via a stimulation of glucose uptake.…”

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confidence: 99%

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Apoptosis is regulated by the rate of glucose transport in an interleukin 3 dependent cell line.

Kan¹,

Baldwin²,

Whetton³

1994

The Journal of Experimental Medicine

110

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In the absence of a survival stimulus, the interleukin 3 (IL-3)-dependent IC.DP cell line undergoes a process termed programmed cell death or apoptosis. Survival can be induced by IL-3, which can also stimulate proliferation of IC.DP cells. IC.DP cells have been stably transfected with the p160v-abl protein tyrosine kinase, activation of the kinase at the permissive temperature permits cell survival in the absence of IL-3 by suppression of apoptosis, although the growth factor is still required for proliferation. Both IL-3 and activation of the v-ABL tyrosine kinase stimulated glucose transport, which may in part be due to a translocation of transporters to the cell surface. Inhibition of glucose uptake markedly increased the rate of apoptosis in these cells, an effect that could be reversed by the provision of alternative energy sources such as glutamine. Growth factor- or oncogene-mediated increases in glucose uptake may therefore represent an important regulatory point in the suppression of apoptosis.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Apoptosis is regulated by the rate of glucose transport in an interleukin 3 dependent cell line.

Kan¹,

Baldwin²,

Whetton³

1994

The Journal of Experimental Medicine

110

View full text Add to dashboard Cite

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Further characterisation of the in situ terminal deoxynucleotidyl transferase (TdT) assay for the flow cytometric analysis of apoptosis in drug resistant and drug sensitive leukaemic cells

et al. 1995

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Apoptosis, originally defined by specific morphological changes, is characterised biochemically by non-random cleavage of DNA. Depending on cell type, this DNA cleavage proceeds from 300 and 5Okbp fragments prior to, concomitantly with, or in the absence of l80bp integer fragmentation. Incorporation into fragmented DNA of biotin-labelled nucleotides by terminal deoxynucleotidyl transferase (TdT) has recently become a standard flow cytometric assay for the idenacation and quantitation of apoptosis. Nucleotide incorportion is visualized using avidin-tagged fluorescein isothiocyanate (FITC) (Gorczyca et al.: Cancer Res 53:1945-1951; Jonker et al.: Cytometry (Suppl13)Abstr 99A, 1993). Here, we charaderise this assay further in three different haemopoietic cell lines. Drug-induced DNA damage is not i d e n a e d by the TdT assay unless it is coupled to the apoptotic response.This was demonstrated using cells in which activation of the oncogenic Abelson-encoded protein tyrosine kinase suppressed drug-induced apoptosis, but did not inhibit drug-induced DNA damage (by melphalan, hydroxyurea, or etoposide). Furthermore, the TdT assay identifies DNA fragments formed during apoptosis induced by etoposide and N-methylformamide in HL60 and MOLT-4 cells, including those high molecular weight DNA fragments formed in MOLT-4 cells which were not further cleaved to 180-2OObp integer fragments. Our results support the use of flow cytometry and the TdT assay to reliably measure apoptotic cells in heterogeneous cell samples. 0 1995 Wiley-Liss, Inc.Key terms: Apoptosis, drug resistance, Abelson tyrosine kinase; DNA damage, non-random DNA fragmentation, TdT assay Apoptosis is now widely recognised as the predominant mode of cell death induced by anticancer agents in sensitive tumour cells (25). An increasing body of evidence demonstrates that apoptotic cell death is actively regulated ( 17,52,54); several proteins suppress apoptosis downstream of drug-induced DNA damage including Bcl-2 (20,38,51), Bcr-Abl (37), and the virally encoded proteins, adenovirus E1B (14) and v-Abl(7). The tumour suppressor gene pS3 is thought to sense DNA strand breaks and to either block cell cycle progression and/or induce apoptotic cell death (8,32,35,36,39). Being an active process, the apoptotic pathway presents opportunities for therapeutic intervention; understanding the molecular mechanisms underlying the induction, and perhaps more importantly, the suppression of apoptosis in tumour cells is now a major thrust of much cancer research.The accurate measurement of apoptosis is problematic; apoptosis in vitro is generally rapid and asynchronous with final non-specific degradation of apoptotic cells (secondary necrosis). The most commonly used, the most certain, and the most laborious method for identification and quantitation of apoptosis is examination of the characteristic changes in nuclear morphology of apoptotic cells by microscopy. However, over the past S years flow cytometric assays for apoptosis have been developed; these (to a large extent) o...

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Deregulation of Signal Transduction Pathways by Oncogenic Retroviruses

Ruscetti

Cmarik²

2010

Retroviruses and Insights Into Cancer

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Cellular signaling events elicited by v-abl associated with growth factor independence in an interleukin-3-dependent cell line

Cited by 29 publications

References 43 publications

Apoptosis is regulated by the rate of glucose transport in an interleukin 3 dependent cell line.

Apoptosis is regulated by the rate of glucose transport in an interleukin 3 dependent cell line.

Further characterisation of the in situ terminal deoxynucleotidyl transferase (TdT) assay for the flow cytometric analysis of apoptosis in drug resistant and drug sensitive leukaemic cells

Deregulation of Signal Transduction Pathways by Oncogenic Retroviruses

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