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Titolo:
CORTICOTROPIN-RELEASING FACTOR RECEPTORS IN HUMAN SMALL-CELL LUNG-CARCINOMA CELLS - RADIOLIGAND BINDING, 2ND-MESSENGER, AND NORTHERN BLOT ANALYSIS DATA
Autore:
DIETERICH KD; GRIGORIADIS DE; DESOUZA EB;
Indirizzi:
NEUROCRINE BIOSCI INC,3050 SCI PK RD SAN DIEGO CA 92121 NEUROCRINE BIOSCI INC SAN DIEGO CA 92121 UNIV CALIF SAN DIEGO,DEPT NEUROSCI SAN DIEGO CA 92037
Titolo Testata:
Endocrinology
fascicolo: 4, volume: 135, anno: 1994,
pagine: 1551 - 1558
SICI:
0013-7227(1994)135:4<1551:CFRIHS>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
ADENYLATE-CYCLASE ACTIVITY; CENTRAL NERVOUS-SYSTEM; FACTOR CRF RECEPTORS; AUTORADIOGRAPHIC LOCALIZATION; ANTERIOR-PITUITARY; NONPEPTIDE ANTAGONISTS; MOLECULAR-WEIGHT; DIVALENT-CATIONS; BETA-ENDORPHIN; MOUSE SPLEEN;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
48
Recensione:
Indirizzi per estratti:
Citazione:
K.D. Dieterich et al., "CORTICOTROPIN-RELEASING FACTOR RECEPTORS IN HUMAN SMALL-CELL LUNG-CARCINOMA CELLS - RADIOLIGAND BINDING, 2ND-MESSENGER, AND NORTHERN BLOT ANALYSIS DATA", Endocrinology, 135(4), 1994, pp. 1551-1558

Abstract

Numerous peptides, growth factors, and receptors have been identifiedin small cell lung carcinoma (SCLC) cells. The present study was designed to examine the radioligand binding, second messenger, and messenger RNA (mRNA) characteristics of CRF receptors in a variety of SCLC lines and to compare their characteristics to CRF receptors in the mousepituitary tumor AtT-20 cells. The human SCLC cell lines NCI-H69, H82,H146, H209, H345, H446, and H510A and control AtT-20 cells all demonstrated specific [I-125]Tyr degrees-ovine CRF ([I-125]oCRF) binding, which was linear with increasing protein concentrations, saturable, reversible, and of high affinity. NCI-H82 cells showed the highest level of specific [I-125]oCRF binding (similar to 60% of the total binding). Scatchard analysis revealed a single homogeneous class of binding sites in NCI-H82 and AtT-20 cells, with K-d values of 263 +/- 48 and 285 +/- 75 pM, respectively, and binding capacities of 74 +/- 7 and 70 +/- 13 fmol/mg protein, respectively. [I-125]oCRF-binding sites on NCI-H82and AtT-20 cells had comparable pharmacological characteristics with the following rank order of inhibitory potencies: rat/human CRF similar to ovine CRF similar to bovine CRF > alpha-helical oCRF-(9-41) > bovine CRF-(1-41)OH much greater than vasoactive intestinal peptide, secretin, GH-releasing hormone. [I-125]oCRF binding in the cell lines was inhibited by guanine nucleotides, suggesting a coupling of receptors to guanine nucleotide-binding proteins. The functional nature of the CRF receptor was demonstrated in second messenger studies in which rat/human CRF stimulated cAMP production in NCI-H82 and AtT-20 cells with comparable EC(50) values of about 3 nM; the percent stimulation over basal activity was significantly higher in NCI-H82 cells (similar to 30-fold increase) than in AtT-20 cells (similar to 12-fold increase). Northern blot analysis of total RNA revealed the presence of a 2.6-kilobase mRNA band in NCI-H82 cells corresponding to the recently cloned human CRF receptor. In summary, the data demonstrate the presence of CRF receptors in SCLC cell lines with kinetic, pharmacological, second messenger, and mRNA characteristics comparable to those in pituitary and brain and suggest a possible role for CRF as a regulatory peptide in human SCLC.

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Documento generato il 28/11/20 alle ore 00:42:21