The distinct role of guanine nucleotide exchange factor Vav1 in Bcl-2 transcription and apoptosis inhibition in Jurkat leukemia T cells.

ABSTRACT:

Aim

To investigate a novel function of proto-oncogene Vav1 in the apoptosis of human leukemia Jurkat cells.

Methods

Jurkat cells, Jurkat-derived vav1-null cells (J.Vav1) and Vav1-reconstituted J.WT cells were treated with a Fas agonist antibody, IgM clone CH11. Apoptosis was determined using propidium iodide (PI) staining, Annexin-V staining, DNA fragmentation, cleavage of caspase 3/caspase 8, and poly (ADP-ribose) polymerase (PARP). Mitochondria transmembrane potential (ΔΨ(m)) was measured using DiOC(6)(3) staining. Transcription and expression of the Bcl-2 family of proteins were evaluated using semi-quantitative RT-PCR and Western blot, respectively. Bcl-2 promoter activity was analyzed using luciferase reporter assays.

Results

Cells lacking Vav1 were more sensitiv

SUBMITTER: Yin J 

PROVIDER: S-EPMC4003318 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature