title: High glucose promotes TGF-β1 production by inducing FOS expression in human peritoneal mesothelial cells.
abstract: BACKGROUND
  High glucose (HG) induces production of transforming growth factor-beta1 (TGF-β1), but the mechanism remains elusive. The aim of this study was to determine the gene(s) involved in HG-induced TGF-β1 production in human peritoneal mesothelial cells (HPMCs).
  
  
  METHODS
  Microarray analysis was performed following a 3-h preincubation of HPMCs in 4 or 0.1% glucose medium. Transcriptional genes were selected using Gene Ontology analysis for biological processes, including regulation of transcription and DNA-dependent. The effects of small interfering RNA (siRNA) treatments on the up-regulation of TGF-β1 mRNA were assessed by quantitative real-time polymerase chain reaction (qPCR). Finally, enzyme-linked immunosorbent assay (ELISA) was performed to determine which gene(s) contribute to the production of TGF-β1 protein in the medium.
  
  
  RESULTS
  Microarray analysis revealed that the expression of 51 genes increased by more than 3-fold. Gene ontology analysis identified 13 genes for further study. qPCR confirmed mRNA amplification for 9 of the 13 genes. Furthermore, HG-induced up-regulation of TGF-β1 mRNA was attenuated by the siRNA of 4 genes: MDS1 and EVI1 complex locus (MECOM), FBJ murine osteosarcoma viral oncogene homolog B (FOSB), FBJ murine osteosarcoma viral oncogene homolog (FOS) and activating transcription factor 3 (ATF3). ELISA showed that siRNA treatment of FOS, but not MECOM, FOSB or ATF3, suppressed the increase of TGF-β1 protein in the medium.
  
  
  CONCLUSIONS
  FOS is a downstream effector of HG stimulation in HPMCs that contributes to TGF-β1 production, suggesting that blocking FOS expression may be a therapeutic target for peritoneal fibrosis.
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Kokoroishi
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan. ayumu@hiroshima-u.ac.jp.|Department of Regeneration and Medicine, Medical Center for Translational and Clinical Research, Hiroshima University Hospital, 1-2-3 Kasumi Minami-Ku, Hiroshima, 734-8553, Japan. ayumu@hiroshima-u.ac.jp.
authors.forename: Ayumu
authors.initials: A
authors.lastname: Nakashima
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan. sdoi@hiroshima-u.ac.jp.|Department of Blood Purification, Hiroshima University Hospital, 1-2-3 Kasumi Minami-Ku, Hiroshima, 734-8553, Japan. sdoi@hiroshima-u.ac.jp.
authors.forename: Shigehiro
authors.initials: S
authors.lastname: Doi
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan.
authors.forename: Toshinori
authors.initials: T
authors.lastname: Ueno
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Doi
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Yokoyama
authors.affiliation: Department of Dental and Medical Biochemistry, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Kiyomasa
authors.initials: K
authors.lastname: Honda
authors.affiliation: Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan.
authors.forename: Masami
authors.initials: M
authors.lastname: Kanawa
authors.affiliation: Department of Dental and Medical Biochemistry, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Kato
authors.affiliation: Department of Molecular and Internal Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Nobuoki
authors.initials: N
authors.lastname: Kohno
authors.affiliation: Department of Nephrology, Hiroshima University Hospital, Hiroshima, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Masaki
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis; D010537:Peritoneum / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D016760:Proto-Oncogene Proteins c-fos; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D005947:Glucose
keywords: FOS; High glucose; Human peritoneal mesothelial cells; Microarray; TGF-β1