title: Hic-5 contributes to epithelial-mesenchymal transformation through a RhoA/ROCK-dependent pathway.
abstract: Epithelial-mesenchymal transformation (EMT) in response to TGFbeta1 is a coordinated process of tissue morphogenesis that occurs during embryonic development as well as during certain pathologic events including kidney tubulointerstitial fibrosis. It is characterized by the disassembly of cell-cell junctions and dramatic alterations in the actin cytoskeleton that facilitates cell-matrix adhesion and stimulates migration. The focal adhesion adapter protein, Hic-5, has previously been reported to be upregulated during TGFbeta1-induced EMT in mouse mammary epithelial cells and the current study recapitulates this result in both mouse kidney proximal tubule epithelial, MCT, cells and human mammary epithelial, MCF10A, cells. To evaluate a causative role for Hic-5 in EMT, Hic-5 RNA interference (siRNA) was used to prevent Hic-5 expression in response to TGFbeta1 stimulation and was shown to suppress cell migration and actin stress fiber formation. It also resulted in the retention of a robust epithelial cell morphology characterized by elevated E-cadherin protein expression and well-organized adherens junctions. In addition, Hic-5 siRNA treatment led to the suppression of TGFbeta1 induction of RhoA activation. In contrast, forced expression of Hic-5 led to the formation of ROCK-dependent actin stress fibers. Furthermore, the induction of Hic-5 expression in response to TGFbeta1 was shown to be a RhoA/ROCK I-dependent process. Together, these data implicate Hic-5 as a key regulator of EMT and suggest that RhoA stimulated Hic-5 expression in response to TGFbeta1 may be functioning in a feed forward mechanism whereby Hic-5 maintains the mesenchymal phenotype through sustained RhoA activation and signaling.
authors.affiliation: Department of Cell and Developmental Biology, State University of New York Upstate Medical University, Syracuse, New York 13210, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Tumbarello
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Turner
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D005355:Fibrosis; D022001:Focal Adhesions / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D007680:Kidney Neoplasms / Q000378:metabolism / Q000473:pathology; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000196:embryology; D060588:LIM Domain Proteins; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D022502:Stress Fibers / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D015854:Up-Regulation / Q000502:physiology; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D034741:RNA, Small Interfering; C090090:Tgfb1i1 protein, mouse; D053773:Transforming Growth Factor beta1; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein