P21WAF/CIP1/SDI1 is upregulated due to increased mRNA stability during hydroxyurea-induced senescence of human fibroblasts

Abstract Hydoxyurea induces senescence-like growth arrest in normal human fibroblasts. p21 WAF/CIP1/SDI1 , a cyclin dependent kinase inhibitor, was found to be upregulated during this growth arrest. Levels of p21 WAF/CIP1/SDI1 protein and mRNA were increased nine-fold by hydroxyurea in these cells. In order to determine whether p21 WAF/CIP1/SDI1 mRNA is increased by hydroxyurea at the transcriptional level, human fibroblast cells were transfected with reporter constructs containing a p21 WAF/CIP1/SDI1 promoter fragment and then treated with hydroxyurea. The luciferase activities in the reporter-transfected fibroblast cells were not increased by hydroxyurea, indicating that p21 WAF/CIP1/SDI1 transcription was not elevated by hydroxyurea. The half-life of the p21 WAF/CIP1/SDI1 mRNA was increased by 2.5-fold but that of p21 WAF/CIP1/SDI1 protein was not. Our results suggest that increased mRNA stability is the major mechanism of p21 WAF/CIP1/SDI1 elevation in the hydroxyurea-induced growth arrest of human fibroblasts.