p53-independent regulation of cyclin B1 in normal human fibroblasts during UV-induced G2-arrest.
Recently we demonstrated, using normal human fibroblasts (NHFs), that UVc radiation induces a G2/M arrest which was even more pronounced when p53 expression was inhibited. So, the aim of this study was to evaluate in NHFs the relationship between UV-induced G2/M arrest and cyclin B1 regulation and to investigate if p53 could contribute to the cyclin B1 regulation in these conditions. Following exposure of asynchronous NHFs to UV light, we showed that the induced G2/M arrest was accompanied by a dose-dependent down-regulation of cyclin B1 mRNA as evaluated by RT-PCR. Concomitantly, using flow cytometric analysis, we observed a strong accumulation of cyclin B1 protein which was correlated to the apparition of the G2/M arrest. In order to study the contribution of p53 to the cyclin B1 accumulation in response to UV exposure, we inhibited p53 induction using p53 antisense oligonucleotides. We found that the inhibition of p53 protein induction after UV exposure had no effect on the level of cyclin B1 mRNA. Moreover, although inhibition of p53 protein induction increased the number of the cells in the G2-M phase, the mean content of cyclin B1 protein was not augmented in these cells. These results indicate clearly that the induction of p53 protein following UV exposure does not regulate the level of cyclin B1 mRNA or protein in normal cells.
Vincent
F
F
Laboratoire d'Oncologie Moléculaire, Universitaires de Strasbourg, France.
Deplanque
G
G
Ceraline
J
J
Duclos
B
B
Bergerat
J P
JP
D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000528:radiation effects; D019926:Cyclin B / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D056744:Cyclin B1; D005347:Fibroblasts / Q000737:chemistry* / Q000502:physiology / Q000528:radiation effects; D005434:Flow Cytometry; D016195:G2 Phase / Q000528:radiation effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007231:Infant, Newborn; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000494:pharmacology* / Q000502:physiology; D014466:Ultraviolet Rays
C534699:CCNB1 protein, human; D019926:Cyclin B; D056744:Cyclin B1; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53