Enhanced sensitivity to hydrogen peroxide-induced apoptosis in Evi1 transformed Rat1 fibroblasts due to repression of carbonic anhydrase III

P. Roy, E. Reavey, M. Rayne, S. Roy, M. Abed El Baky, Yuji Ishii, C. Bartholomew

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

EVI1 is a nuclear zinc finger protein essential to normal development, which participates in acute myeloid leukaemia progression and transforms Rat1 fibroblasts. In this study we show that enforced expression of Evi1 in Rat1 fibroblasts protects from paclitaxel-induced apoptosis, consistent with previously published studies. Surprisingly, however, these cells show increased sensitivity to hydrogen peroxide (H2O2)-induced apoptosis, demonstrated by elevated caspase 3 catalytic activity. This effect is caused by a reduction in carbonic anhydrase III (caIII) production. caIII transcripts are repressed by 92-97% by Evi1 expression, accompanied by a similar reduction in caIII protein. Reporter assays with the rat caIII gene promoter show repressed activity, demonstrating that Evi1 either directly or indirectly modulates transcription of this gene in Rat1 cells. Targeted knockdown of caIII alone, with Dicer-substrate short inhibitory RNAs, also increases the sensitivity of Rat1 fibroblasts to H2O2, which occurs in the absence of any other changes mediated by Evi1 expression. Enforced expression of caIII in Evi1-expressing Rat1 cells reverts the phenotype, restoring H2O 2 resistance. Together these data show that Evi1 represses transcription of caIII gene expression, leading to increased sensitivity to H2O2-induced apoptosis in Rat1 cells and might suggest the basis for the development of a novel therapeutic strategy for the treatment of leukaemias and solid tumours where EVI1 is overexpressed.

Original languageEnglish
Pages (from-to)441-452
Number of pages12
JournalFEBS Journal
Volume277
Issue number2
DOIs
Publication statusPublished - Jan 1 2010

Fingerprint

Carbonic Anhydrase III
Fibroblasts
Hydrogen Peroxide
Apoptosis
Transcription
Genes
Zinc Fingers
Paclitaxel
Acute Myeloid Leukemia
Gene expression
Caspase 3
Zinc
Rats
Tumors
Assays
Catalyst activity
Leukemia
Proteins
RNA
Phenotype

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Enhanced sensitivity to hydrogen peroxide-induced apoptosis in Evi1 transformed Rat1 fibroblasts due to repression of carbonic anhydrase III. / Roy, P.; Reavey, E.; Rayne, M.; Roy, S.; Abed El Baky, M.; Ishii, Yuji; Bartholomew, C.

In: FEBS Journal, Vol. 277, No. 2, 01.01.2010, p. 441-452.

Research output: Contribution to journalArticle

Roy, P. ; Reavey, E. ; Rayne, M. ; Roy, S. ; Abed El Baky, M. ; Ishii, Yuji ; Bartholomew, C. / Enhanced sensitivity to hydrogen peroxide-induced apoptosis in Evi1 transformed Rat1 fibroblasts due to repression of carbonic anhydrase III. In: FEBS Journal. 2010 ; Vol. 277, No. 2. pp. 441-452.
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