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  1. Article: Paracrine calcitonin in prostate cancer is linked to CD44 variant expression and invasion.

    Iczkowski, Kenneth A / Omara-Opyene, A Levi / Kulkarni, Trupti R / Pansara, Megha / Shah, Girish V

    Anticancer research

    2005  Volume 25, Issue 3B, Page(s) 2075–2083

    Abstract: Background: Calcitonin (CT) exerts an autocrine/paracrine influence on prostatic tumor invasion through coupling to transduction protein Gsalpha. Cell adhesion glycoprotein CD44 variant v7-v10 also faciliates invasion, but its modulation by the CT- ... ...

    Abstract Background: Calcitonin (CT) exerts an autocrine/paracrine influence on prostatic tumor invasion through coupling to transduction protein Gsalpha. Cell adhesion glycoprotein CD44 variant v7-v10 also faciliates invasion, but its modulation by the CT-Gsalpha system was unexplored.
    Materials and methods: LnCaP, PC-3 and metastasis-derived PC-3M cell lines were studied, including cells modified therefrom: Gsalpha-QL, expressing mutant constitutively active Gsalpha protein, and CT+, overexpressing CT. CD44 variant expression was evaluated in vivo after orthotopic implantion into nude mice, and in vitro by real-time RT-PCR and Western blotting.
    Results: Both mRNA and protein levels of the CD44 variant were minimal in PC-3M tumor implants, but elevated in Gsalpha-QL. Exogenous CT stimulated invasion into Matrigel strongly in LnCaP and CT+, and less in PC-3 and Gsalpha-QL. By Western blot analysis, untreated Gsalpha-QL and CT+ cells overexpressed CD44 variant compared with LnCaP or PC-3. By quantitative RT-PCR, exogenous CT dose-dependently increased CD44 variant mRNA to seven-fold. Pharmacologic agents that stimulated or inhibited Gsalpha activity or stimulated adenylyl cyclase produced proportionate dose-dependent effects on both CD44 variant expression and Matrigel invasion.
    Conclusion: This paracrine factor, acting though cyclic AMP, regulates the expression of CD44v7-10, which modulates the tumor phenotype.
    MeSH term(s) Adenylyl Cyclases/metabolism ; Animals ; Calcitonin/metabolism ; Calcitonin/pharmacology ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; GTP-Binding Protein alpha Subunits, Gs/biosynthesis ; GTP-Binding Protein alpha Subunits, Gs/metabolism ; Guanylyl Imidodiphosphate/pharmacology ; Humans ; Hyaluronan Receptors/biosynthesis ; Hyaluronan Receptors/genetics ; In Situ Hybridization ; Male ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Prostatic Neoplasms/genetics ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/pathology ; Protein Isoforms ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics
    Chemical Substances CD44v9 antigen ; Hyaluronan Receptors ; Protein Isoforms ; RNA, Messenger ; Guanylyl Imidodiphosphate (34273-04-6) ; Calcitonin (9007-12-9) ; GTP-Binding Protein alpha Subunits, Gs (EC 3.6.5.1) ; Adenylyl Cyclases (EC 4.6.1.1)
    Language English
    Publishing date 2005-05
    Publishing country Greece
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 604549-2
    ISSN 1791-7530 ; 0250-7005
    ISSN (online) 1791-7530
    ISSN 0250-7005
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1642: Show issues Location:
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    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
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  2. Article: The loop C region of the murine 5-HT3A receptor contributes to the differential actions of 5-hydroxytryptamine and m-chlorophenylbiguanide.

    Suryanarayanan, Asha / Joshi, Prasad R / Bikádi, Zsolt / Mani, Muthalagi / Kulkarni, Trupti R / Gaines, Chandra / Schulte, Marvin K

    Biochemistry

    2005  Volume 44, Issue 25, Page(s) 9140–9149

    Abstract: Sequence and predicted structural similarities between members of the Cys loop superfamily of ligand-gated ion channel receptors and the acetylcholine binding protein (AChBP) suggest that the ligand-binding site is formed by six loops that intersect at ... ...

    Abstract Sequence and predicted structural similarities between members of the Cys loop superfamily of ligand-gated ion channel receptors and the acetylcholine binding protein (AChBP) suggest that the ligand-binding site is formed by six loops that intersect at subunit interfaces. We employed site-directed mutagenesis to investigate the role of amino acids from the loop C region of the murine 5-HT(3AS)R in interacting with two structurally different agonists, serotonin (5-HT) and m-chlorophenylbiguanide (mCPBG). Mutant receptors were evaluated using radioligand binding, two-electrode voltage clamp, and immunofluorescence studies. Electrophysiological assays were employed to identify changes in response characteristics and relative efficacies of mCPBG and the partial agonist, 2-methyl 5-HT (2-Me5-HT). We have also constructed novel 5-HT and mCPBG docked models of the receptor binding site based on homology models of the AChBP. Both ligand-docked models correlate well with results from mutagenesis and electrophysiological assays. Four key amino acids were identified as being important to ligand binding and/or gating of the receptor. Among these, I228 and D229 are specific for effects mediated by 5-HT compared to mCPBG, indicating a differential interaction of these ligands with loop C. Residues F226 and Y234 are important for both 5-HT and mCPBG interactions. Mutations at F226, I228, and Y234 also altered the relative efficacies of agonists, suggesting a role in the gating mechanism.
    MeSH term(s) Amino Acid Sequence ; Animals ; Biguanides/chemistry ; Biguanides/metabolism ; Cell Line ; Electrophysiology ; Humans ; Methylation ; Mice ; Models, Molecular ; Mutation/genetics ; Oocytes/metabolism ; Patch-Clamp Techniques ; Protein Structure, Tertiary ; Radioligand Assay ; Receptors, Serotonin, 5-HT3/chemistry ; Receptors, Serotonin, 5-HT3/genetics ; Receptors, Serotonin, 5-HT3/metabolism ; Sequence Alignment ; Serotonin/chemistry ; Serotonin/metabolism ; Serotonin 5-HT3 Receptor Agonists ; Serotonin Receptor Agonists/chemistry ; Serotonin Receptor Agonists/metabolism ; Xenopus laevis
    Chemical Substances Biguanides ; Receptors, Serotonin, 5-HT3 ; Serotonin 5-HT3 Receptor Agonists ; Serotonin Receptor Agonists ; Serotonin (333DO1RDJY) ; 1-(3-chlorophenyl)biguanide (910A4X901V)
    Language English
    Publishing date 2005-06-28
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 1108-3
    ISSN 1520-4995 ; 0006-2960
    ISSN (online) 1520-4995
    ISSN 0006-2960
    DOI 10.1021/bi050661e
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 217: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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