Cyclic AMP regulation of pro-vasoactive intestinal polypeptide/PHM-27 synthesis in human neuroblastoma cells.

Abstract

The mechanism of N6,O2'-dibutyryl adenosine 3':5'-monophosphate (Bt2cAMP) induction of pro-vasoactive intestinal polypeptide (VIP)/PHM-27 biosynthesis was investigated in human neuroblastoma cells in culture. When neuroblastoma cells were grown for 48 h in the presence of 1 mM Bt2cAMP, the synthesis of pro-VIP/PHM-27 was stimulated 11-fold. The amount of prepro-VIP/PHM-27 mRNA determined both by hybridization with cloned prepro-VIP/PHM-27 cDNA and a reticulocyte cell-free translation assay was also increased 11-fold in the Bt2cAMP-induced cells. Transcription of prepro-VIP/PHM-27 mRNA in isolated nuclei was observed in induced cells, but not in uninduced cells. Blot hybridization with prepro-VIP/PHM-27 cDNA of total nuclear RNA isolated from neuroblastoma cells revealed an RNA species corresponding to mature prepro-VIP/PHM-27 mRNA, and the amount of the RNA was markedly increased in the induced cells. The quantity of VIP/PHM-27 gene in the DNA of neuroblastoma cells was analyzed after hydrolysis with a restriction endonuclease, EcoRI. However, VIP/PHM-27 gene was not amplified in the induced cells. These results indicate that Bt2cAMP-induced pro-VIP/PHM-27 synthesis is achieved by enhancing the transcription rate of prepro-VIP/PHM-27 mRNA.