Somatostatin production by a human medullary thyroid carcinoma cell line.
Somatostatin (SRIF, SRIF-14) is a known product of the normal and malignant parafollicular cell of the thyroid. In this report we characterize SRIF production by the TT cells, a line of transformed calcitonin-producing cells derived from a human medullary thyroid carcinoma. The cells were found to contain (5-12 ng/10(6) cells) and secrete (3-10 ng/10(6) cells X 48 h) immunoreactive SRIF. Molecular sieve chromatography of cell extracts under denaturing conditions showed a major peak with a mol wt slightly larger than 12,700, probably representing pro-SRIF and a second peak which coeluted with SRIF; in one gel chromatogram a very small peak was also noted which coeluted with SRIF-28, but represented less than 0.4% of the total immunoreactive SRIF. Short term secretion of calcitonin and SRIF was stimulated by calcium in vitro (0.5-4 mM) in a dose-related manner. mRNA isolated from the TT cells hybridized to a specific bovine fetal pancreatic SRIF DNA (BFPS-2); there was no hybridization to identical amounts of mRNA from the atT-20/D16, 3T3, or RINC5F cell lines. In vitro translation of the TT cell mRNA followed by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the product revealed a single protein band of approximately 13,000 daltons. It was completely abolished when the immunoprecipitation was performed in the presence of excess unlabeled SRIF. Northern transfer of TT cell cytoplasmic RNA and hybridization with FBPS-2 cDNA showed a single hybridizing band with an apparent size of approximately 750 nucleotides. Our observations demonstrate the production of SRIF by a continuous line of human medullary thyroid carcinoma cells and provide a model for studying the biosynthesis and secretion of SRIF in the parafollicular cell.
Gagel
R F
RF
Palmer
W N
WN
Leonhart
K
K
Chan
L
L
Leong
S S
SS
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D002850:Chromatography, Gel; D004247:DNA / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D006801:Humans; D009693:Nucleic Acid Hybridization; D011188:Potassium / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D011863:Radioimmunoassay; D013004:Somatostatin / Q000096:biosynthesis* / Q000235:genetics; D013964:Thyroid Neoplasms / Q000378:metabolism*
D012333:RNA, Messenger; D013004:Somatostatin; D004247:DNA; D011188:Potassium; D002118:Calcium