Procedure for the quantitation of Gadd45 expression levels in clonal hematopoietic progenitor cells by competitive polymerase chain reaction

Objective: The growth arrest and DNA damage-inducible (gadd) genes represen t a family of stress-inducible genes that are coordinately regulated at tra nscriptional level. Gadd45, in particular, has been linked to a p53-depende nt inducible network required for regulated transition from G(1) to S phase of cell cycle following genotoxic insult and growth arrest treatments and has seemingly a pivotal role in DNA repair. Design and methods: Here we show that competitive polymerase chain reaction (PCR) is an adequate method to quantitate gadd45 expression levels in hema topoietic progenitor cell line 32D, whose constitutive gene expression is v ery low. Results: The sensitivity and reproducibility of our strategy support its us efulness for clinical purposes, to assess the DNA repair capacity of highly purified early myeloid progenitors, whose failure may be responsible for e ither short-term chemotherapy side effects (bone marrow hypoplasia and peri pheral blood cytopenia) or longterm consequences of antiblastic drugs (leuk emia and myelodysplasia). Copyright (C) 1999 The Canadian Society of Clinic al Chemists.