title: Laminin-1 and alpha6beta1 integrin regulate acinar morphogenesis of normal and malignant human prostate epithelial cells.
abstract: BACKGROUND
  Cell-matrix interactions via integrin receptors are critical for acinar morphogenesis. The non-tumorigenic, human prostate epithelial cell line RWPE-1 was used in a three-dimensional (3D) cell culture model to identify the matrix protein and its integrin receptor required for acinar morphogenesis.
  
  
  METHODS
  3D cultures, immunostaining, confocal microscopy, and Western blot analysis were used to examine acinar formation on matrix proteins and to determine integrin receptor expression.
  
  
  RESULTS
  RWPE-1 cells differentiate into acini of polarized cells with a distinct lumen in 3D Matrigel culture. In contrast, the malignant WPE1-NB26 prostate epithelial cells form solid cell masses. In 3D gels of laminin-1, type IV collagen, or fibronectin, RWPE-1 cells form acini only in laminin-1. Anti-laminin-1 antibody reduces acinar formation in a dose-dependent manner. Polarized RWPE-1 cells showed basal expression of alpha6 and beta1 integrin subunits. Blocking antibodies to alpha6 or beta1 reduced acinar formation to 9 and 6 percent of control, respectively. The beta1 integrin colocalized with focal adhesion kinase (FAK). Inhibition of extracellular signal-regulated kinase kinase activity significantly reduced acinar formation to 38 percent of control, suggesting that beta1 integrin-mediated signal transduction may be regulated through a FAK pathway.
  
  
  CONCLUSIONS
  While basal expression of alpha6beta1 integrin in RWPE-1 cells correlates with their ability to polarize and form acini, a decrease or loss of alpha6, and diffused beta1 expression in WPE1-NB26 cells correlates with loss of acinar-forming ability. Results show that laminin-1 and a functional alpha6beta1 integrin receptor are required for acinar morphogenesis. This novel 3D cell culture model is useful for elucidating regulation of acinar morphogenesis and its loss during prostate carcinogenesis.
authors.affiliation: Department of Medicine, Michigan State University, East Lansing, MI 48824-1312, USA.
authors.forename: D
authors.initials: D
authors.lastname: Bello-DeOcampo
authors.forename: H K
authors.initials: HK
authors.lastname: Kleinman
authors.forename: N D
authors.initials: ND
authors.lastname: Deocampo
authors.forename: M M
authors.initials: MM
authors.lastname: Webber
mesh_terms: D000906:Antibodies / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D003094:Collagen; D004338:Drug Combinations; D004789:Enzyme Activation; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006801:Humans; D039121:Integrin alpha6beta1; D016023:Integrins / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology*; D007797:Laminin / Q000378:metabolism / Q000502:physiology*; D048369:MAP Kinase Kinase 1; D020935:MAP Kinase Signaling System / Q000502:physiology; D008297:Male; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors; D011467:Prostate / Q000166:cytology* / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D011505:Protein-Tyrosine Kinases / Q000378:metabolism / Q000502:physiology; D011509:Proteoglycans; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000096:biosynthesis
chemical_list: D000906:Antibodies; D004338:Drug Combinations; D039121:Integrin alpha6beta1; D016023:Integrins; D007797:Laminin; D011509:Proteoglycans; D014746:Vimentin; C121346:laminin 1; C059485:matrigel; D003094:Collagen; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human; D017346:Protein Serine-Threonine Kinases; D048369:MAP Kinase Kinase 1; C482125:MAP2K1 protein, human; D020929:Mitogen-Activated Protein Kinase Kinases