Detection of multiple hormones and their mRNAs in human neuroblastoma cell line NB-1 using in situ hybridization, immunocytochemistry and radioimmunoassay.
The production and secretion of multiple peptide hormones and tyrosine hydroxylase by the human neuroblastoma cell line NB-1 and the effects of dibutyryl cAMP (Bt2cAMP) and phorbol esters such as 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on them were investigated. The presence of messenger RNAs (mRNAs) of vasoactive intestinal peptide (VIP)/peptide histidine methionine (PHM), preprotachykinin, and tyrosine hydroxylase was detectable in the cytoplasm of cultured NB-1 cells by in situ hybridization. Treatment with Bt2cAMP and TPA markedly increased the number of cells immunoreactive to VIP, PHM, neuropeptide Y, Met-enkephalin, substance P and tyrosine hydroxylase and also the contents of VIP and Met-enkephalin in the culture medium. Bt2cAMP and TPA induced morphological changes characteristic of endocrine differentiation, such as an increase in neuroendocrine granules and the development of rough endoplasmic reticulum and Golgi apparatus. The results indicated that treatment with Bt2cAMP and TPA induces the expression of multiple genes of peptide hormone and tyrosine hydroxylase and increases hormone production and secretion through morphological changes into endocrine cells.
D003994:Bucladesine / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000648:ultrastructure; D004744:Enkephalin, Methionine / Q000032:analysis; D006728:Hormones / Q000032:analysis* / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008854:Microscopy, Electron; D009447:Neuroblastoma / Q000737:chemistry* / Q000648:ultrastructure; D009478:Neuropeptide Y / Q000032:analysis; D010451:Peptide PHI / Q000032:analysis / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis*; D011863:Radioimmunoassay; D013373:Substance P / Q000032:analysis; D015320:Tachykinins / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000235:genetics; D014660:Vasoactive Intestinal Peptide / Q000032:analysis / Q000235:genetics
D006728:Hormones; D009478:Neuropeptide Y; D010451:Peptide PHI; D011498:Protein Precursors; D012333:RNA, Messenger; D015320:Tachykinins; C044206:preprotachykinin; D013373:Substance P; D014660:Vasoactive Intestinal Peptide; D004744:Enkephalin, Methionine; D003994:Bucladesine; D014446:Tyrosine 3-Monooxygenase; D013755:Tetradecanoylphorbol Acetate