ANTISENSE P53 PROVOKES CHANGES IN HELA-CELL GROWTH AND MORPHOLOGY

Human cervical carcinoma HeLa cells, containing the wild-type p53 prot ein, were stably transfected with a p53 antisense encoding plasmid, Di fferences in the p53 content in extracts from the p53 antisense transf ected HeLa clones compared to the parental HeLa cells were demonstrate d by protein binding to the p53 consensus oligonucleotide and in trans activation assays, Striking morphological alterations were observed in several HeLa clones stably transfected with the p53 antisense encodin g plasmid. Giant multinucleated cells appeared several passages after transfection in four p53 antisense transfected HeLa clones derived fro m three independent transfection experiments, and were not observed in parental HeLa cells, or in p53 sense or vector-transfected HeLa clone s, A strong growth inhibitory effect was observed for the HeLa clones stably transfected with p53 antisense encoding plasmid, Reconstitution experiments with HeLa cells treated with a short p53 antisense oligon ucleotide gave similar results: growth inhibition and giant cells. The se giant multinucleated cells were negative for [H-3]thymidine incorpo ration and displayed dispersed nuclear staining for proliferating cell nuclear antigen, indicating the absence of DNA replication.