title: Mycophenolic acid reverses TGF beta-induced cell motility, collagen matrix contraction and cell morphology in vitro.
abstract: The aim of this study was to elucidate functional and molecular effects of mycophenolic acid (MPA) on non-lymphatic, kidney epithelial cells treated with transforming growth factor (TGF). MPA effects were studied using HK2 cells incubated with EGF and TGF. The reversibility of these effects was verified using guanosine and 8-aminoguanosine. The following assays were applied: cell proliferation, viability, collagen matrix contraction, scratch wound closure, spindle index, FACS with anti-CD29 and anti-CD326, promoter demethylation of RAS protein activator like 1 (RASAL1), as well as gene expression of RASAL1, integrin 1ß (ITGB1) (CD29) and epithelial cell adhesion molecule (EpCam) (CD326). Cell proliferation was inhibited by increasing concentrations of MPA, whereas neither apoptosis nor cytotoxicity was detected. Stimulation with EGF and/or TGF led to a significant collagen matrix contraction that was successfully inhibited by MPA. In addition, scratch wound closure was inhibited by incubation with TGF alone or with EGF. Under the same conditions, cell morphology (spindle shape) and molecular phenotype (ITGB1(High)EpCam(Low)/ITGB1(Low)EpCam(High)) were both significantly changed, suggesting an epithelial to mesenchymal transformation. Cell morphology and motility, as well as molecular phenotype, were reversible after MPA treatment with TGF transformation in both presence/absence of EGF, thereby suggesting a correlation with the previously described antifibrotic effects of MPA. Dysregulation of TGF signal transduction appears to be related to progression of fibrosis. A TGF-transformed kidney epithelial cell line derived from human proximal tubules was used to study whether the immunosuppressive drug: MPA possesses any functional or molecular antifibrotic effects. Functional and morphological in vitro changes induced by both the TGF and epithelial-growth-factor were reversible by treatment with MPA. An inhibitory effect of MPA on the TGF pathway appears to be responsible for the previously described antifibrotic effects of the MPA in the COL4A3-deficient mouse model of renal fibrosis.
authors.affiliation: Department of Clinical Chemistry, University Medical Center Goettingen, Goettingen, Germany.|Department of Clinical Pharmacology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Darinka Todorova
authors.initials: DT
authors.lastname: Petrova
authors.affiliation: Department of Clinical Chemistry, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Brandhorst
authors.affiliation: Department of Clinical Chemistry, University Medical Center Goettingen, Goettingen, Germany.|Department of Clinical Pharmacology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Koch
authors.affiliation: Department of Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Frank Christian
authors.initials: FC
authors.lastname: Schultze
authors.affiliation: Department of Clinical Chemistry, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Eberle
authors.affiliation: Department of Clinical Chemistry, University Medical Center Goettingen, Goettingen, Germany.|Department of Clinical Pharmacology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Walson
authors.affiliation: Department of Clinical Chemistry, University Medical Center Goettingen, Goettingen, Germany.|Department of Clinical Pharmacology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Oellerich
mesh_terms: D000818:Animals; D001324:Autoantigens; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D024141:Collagen Type IV / Q000172:deficiency*; D004195:Disease Models, Animal; D004815:Epidermal Growth Factor / Q000378:metabolism; D005355:Fibrosis / Q000188:drug therapy; D006801:Humans; D007668:Kidney / Q000187:drug effects* / Q000473:pathology*; D051379:Mice; D009173:Mycophenolic Acid / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D001324:Autoantigens; D024141:Collagen Type IV; D016212:Transforming Growth Factor beta; C057112:type IV collagen alpha3 chain; D004815:Epidermal Growth Factor; D009173:Mycophenolic Acid
keywords: epithelial to mesenchymal transition; growth factors; kidney fibrosis; mycophenolic acid; transformation