title: Are growth factor receptors modulated by metformin in human endometrial stromal cells after stimulation with androgen and insulin?
abstract: OBJECTIVE
  To assess the effect of metformin on gene and protein expression of insulin receptor (IR) and IGF-1 (IGF-1R) receptor in human endometrial stromal cells after stimulation with androgen and insulin.
  
  
  METHODS
  Primary culture of endometrial stromal cells stimulated with estrogen, progesterone with or without androgen or insulin, and treated with metformin for 24 and 48 h, followed by RNA (qRT-PCR) and protein (Western blot) extraction and analysis.
  
  
  RESULTS
  IR gene expression was increased after treatment with insulin (2.9-fold change, p = 0.027) and further after metformin treatment (4.7-fold change, p < 0.001), and in IGF-1R, the group treated with insulin (1.83-fold change) and metformin (1.78-fold change) showed more expression, than control group (p < 0.001). Similarly, IR protein expression was increased after addition of metformin and insulin (249,869 ± 15,878) in relation to the other groups (p < 0.001). Furthermore, cells treated with insulin (153,634 ± 29,123) and androgen plus insulin (162,854 ± 86,258) had a higher IR protein expression compared to control (104,654 ± 5,634) and androgen group (71,595 ± 3,439, (p = 0.045 and 0.021). In groups treated with insulin (127,711 ± 4,591) and androgen plus insulin (151,098 ± 5,194) the protein IGF-1R was increased compared to control (79,355 ± 3,470) and the androgen-only group (79,326 ± 3,114) (p < 0.001).
  
  
  CONCLUSIONS
  Metformin in combination with insulin increased IR protein and gene expressions, while it had no influence on the protein expression of IGF-1R in endometrial stromal cells.
authors.affiliation: Laboratório de Biologia Molecular Endócrina e Tumoral, Departamento de Fisiologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil.
authors.forename: Gustavo Dias
authors.initials: GD
authors.lastname: Ferreira
authors.forename: Ariane
authors.initials: A
authors.lastname: Germeyer
authors.forename: Amanda
authors.initials: A
authors.lastname: de Barros Machado
authors.forename: Tadeu Ludwig
authors.initials: TL
authors.lastname: do Nascimento
authors.forename: Ilma Simoni
authors.initials: IS
authors.lastname: Brum
authors.forename: Thomas
authors.initials: T
authors.lastname: Strowitzki
authors.forename: Helena
authors.initials: H
authors.lastname: von Eye Corleta
authors.forename: Edison
authors.initials: E
authors.lastname: Capp
mesh_terms: D000728:Androgens / Q000494:pharmacology; D015153:Blotting, Western; D002478:Cells, Cultured; D004717:Endometrium / Q000166:cytology* / Q000187:drug effects; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D015870:Gene Expression; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology*; D008687:Metformin / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D011374:Progesterone / Q000494:pharmacology; D017526:Receptor, IGF Type 1 / Q000235:genetics / Q000378:metabolism*; D011972:Receptor, Insulin / Q000235:genetics / Q000378:metabolism*; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism
chemical_list: D000728:Androgens; D004967:Estrogens; D007004:Hypoglycemic Agents; D007328:Insulin; D011374:Progesterone; D008687:Metformin; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin