Document Detail


Cysteine proteinase inhibitor in cultured human medullary thyroid carcinoma cells.
MedLine Citation:
PMID:  1602739     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The TT cell line of human medullary thyroid carcinoma, that retains some of the differentiated functions of thyroid C cells including the synthesis and secretion of calcitonin, was found to contain and release into the culture medium cysteine proteinase inhibitor(s), cystatin(s). The major inhibitor, which is similar to, if not identical with, cystatin C, is constitutively released, or secreted, by TT cells. The rate of secretion of cystatin, quantified by titration of inhibition of papain, was stimulated by dibutyryladenosine 3':5'-cyclic monophosphate, forskolin, the calcium ionophore A 23187, and by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). Neither forskolin nor TPA had, however, an effect on the level of the inhibitor in TT cells. Treatment with n-butyrate strongly inhibited the proliferation of TT cells, and led, in 4 to 7 days, to a doubling of the intracellular concentration of cystatins. Northern blot hybridizations to a 32P-labeled riboprobe complementary to human cystatin C cDNA indicated that cAMP, forskolin, and TPA had no effect on the steady-state levels of cystatin C mRNA. These data indicate that release of cystatin(s) from TT cells is regulated by cAMP-calcium-protein kinase C mechanisms that appear to be similar to those that regulate the secretion of calcitonin from these cells. However, in contrast to the calcitonin gene, the expression of the cystatin C gene in these cells is not regulated by cAMP or TPA. By a combination of acetone fractionation, affinity chromatography on Cm-papain-Sepharose, and gel exclusion chromatography a protein of approximately 14 kilodaltons was isolated from TT cells that reacted with antibodies against human cystatin C, and strongly inhibited papain. Cystain secreted by TT cells also had a molecular weight of 14 kilodaltons, and reacted with anti-human cystatin C antibodies. The physiologic and pathologic roles of cystatins in different cell types remain to be established. The TT cells provide a suitable cell type to study the regulation of the expression of the cystatin gene and the mechanism of cystatin release.
Authors:
T Barka; H van der Noen; S Patil
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Laboratory investigation; a journal of technical methods and pathology     Volume:  66     ISSN:  0023-6837     ISO Abbreviation:  Lab. Invest.     Publication Date:  1992 Jun 
Date Detail:
Created Date:  1992-07-10     Completed Date:  1992-07-10     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0376617     Medline TA:  Lab Invest     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  691-700     Citation Subset:  IM    
Affiliation:
Department of Cell Biology and Anatomy, Mount Sinai School of Medicine, City University of New York, New York.
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MeSH Terms
Descriptor/Qualifier:
Blotting, Northern
Butyric Acid
Butyric Acids / pharmacology
Carcinoma / metabolism*
Cystatin C
Cystatins / genetics,  isolation & purification,  metabolism*
Forskolin / pharmacology
Gene Expression
Humans
Immunoenzyme Techniques
Tetradecanoylphorbol Acetate / pharmacology
Thyroid Neoplasms / metabolism*
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
HL39419/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Butyric Acids; 0/CST3 protein, human; 0/Cystatin C; 0/Cystatins; 107-92-6/Butyric Acid; 16561-29-8/Tetradecanoylphorbol Acetate; 66428-89-5/Forskolin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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