title: INSL-3 is expressed in human hyperplastic and neoplastic thyrocytes.
abstract: The insulin-like hormone INSL-3, also named relaxin-like factor (RLF) or Leydig-derived insulin-like peptide (LEY-IL), is expressed in various reproductive tissues and is regarded a marker of differentiation in human testicular Leydig cells. Recently, we have identified differential expression of human INSL-3 in neoplastic Leydig cells and mammary epithelial cells suggesting an involvement of INSL-3 in tumor biology. Here we have investigated the expression of INSL-3 in human thyroid carcinoma cell lines and in the human thyroid gland which has been shown to express transcripts for the G protein coupled INSL-3 receptor LGR8. When we determined the expression of INSL-3 in eight human thyroid carcinoma cell lines, a novel INSL-3 splice variant containing a 95 bp out-of-frame insertion at the beginning of exon II of the INSL-3 gene was discovered. Treatment of the human anaplastic thyroid carcinoma cell line 8505C with diethylstilbestrol (DES) caused a significant dose-dependent transcriptional down-regulation of INSL-3 and a marked up-regulation of LGR8. Employing in situ hybridization to detect INSL-3 transcripts and specific rabbit antisera against the INSL-3 proteins, both INSL-3 isoforms were detected in patients with Graves' disease (n=10), follicular carcinomas (FTC; n=12), papillary carcinomas (PTC; n=9) and undifferentiated anaplastic carcinomas (UTC; n=15). By contrast, thyrocytes of all 15 benign goiter tissues studied were devoid of both INSL-3 isoforms, mRNA and protein. Our data indicate that INSL-3 hormone is up-regulated in hyperplastic and neoplastic human thyrocytes suggesting that the INSL-3 isoforms may serve as additional markers for hyperplastic and neoplastic human thyrocytes. In the anaplastic thyroid carcinoma cell line 8505C, the regulation of both INSL-3 and LGR8 by estrogen may be the first indication of a novel hormonally responsive, auto-/paracrine INSL-3 LGR8 ligand receptor system active in human thyroid carcinoma cells.
authors.affiliation: Department of General, Visceral and Vascular Surgery, Martin-Luther-University Halle-Wittenberg, 06097 Halle, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Hombach-Klonisch
authors.forename: Cuong
authors.initials: C
authors.lastname: Hoang-Vu
authors.forename: Astrid
authors.initials: A
authors.lastname: Kehlen
authors.forename: Raoul
authors.initials: R
authors.lastname: Hinze
authors.forename: Hans-Jürgen
authors.initials: HJ
authors.lastname: Holzhausen
authors.forename: Ekkehard
authors.initials: E
authors.lastname: Weber
authors.forename: Bernd
authors.initials: B
authors.lastname: Fischer
authors.forename: Henning
authors.initials: H
authors.lastname: Dralle
authors.forename: Thomas
authors.initials: T
authors.lastname: Klonisch
mesh_terms: D017398:Alternative Splicing; D001483:Base Sequence; D017931:DNA Primers; D004054:Diethylstilbestrol / Q000494:pharmacology*; D005091:Exons; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006042:Goiter / Q000235:genetics / Q000473:pathology; D006111:Graves Disease / Q000235:genetics / Q000473:pathology; D006801:Humans; D006965:Hyperplasia; D017403:In Situ Hybridization; D007328:Insulin; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013961:Thyroid Gland / Q000473:pathology*; D013964:Thyroid Neoplasms / Q000235:genetics* / Q000473:pathology; D014158:Transcription, Genetic / Q000187:drug effects*; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D007328:Insulin; C084797:Leydig insulin-like protein; D020033:Protein Isoforms; D011506:Proteins; D004054:Diethylstilbestrol