title: Inhibition of cell proliferation and migration after HTRA1 knockdown in retinal pigment epithelial cells.
abstract: OBJECTIVE
  The purpose of this study was to investigate the role of HtrA serine peptidase 1 (HTRA1) in the proliferation and migration of cells of the human retinal pigment epithelial cell line ARPE-19, and the possible mechanisms involved.
  
  
  METHODS
  ARPE-19 cells were transduced by a recombinant lentiviral vector carrying HTRA1-shRNA to knockdown HTRA1 expression. Subsequent HTRA1 gene and HTRA1 protein levels in these cells and control cells were detected by quantitative real-time PCR and Western blot, respectively. Changes in cell proliferation and migration associated with the inhibition of HTRA1 expression were assessed, as well as changes in the mRNA levels of transforming growth factor beta 1 (TGFB1), bone morphogenetic protein 4 (BMP4), and bone morphogenetic protein 2 (BMP2).
  
  
  RESULTS
  The recombinant lentivirus carrying HTRA1-shRNA was successfully generated, as evidenced by reduced levels of HTRA1 mRNA and HTRA1 protein in ARPE-19 cells. The knockdown of HTRA1 in ARPE-19 cells was associated with reduced cellular proliferation and migration, and increased mRNA levels of TGF-β1, BMP4, and BMP2.
  
  
  CONCLUSIONS
  Silence of the HTRA1 gene was associated with significantly higher levels of TGF-β1, BMP4, and BMP2 mRNA and reduction in the proliferation and migration of ARPE-19 cells.
authors.affiliation: Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology and Visual Science Key Laboratory, No. 1 Dongjiaominxiang, Dongcheng District, Beijing, China, drpeixueting@126.com.
authors.forename: Xueting
authors.initials: X
authors.lastname: Pei
authors.forename: Kai
authors.initials: K
authors.lastname: Ma
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.forename: Ningli
authors.initials: N
authors.lastname: Wang
authors.forename: Ningpu
authors.initials: N
authors.lastname: Liu
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology*; D020868:Gene Silencing / Q000502:physiology*; D005822:Genetic Vectors; D000074289:High-Temperature Requirement A Serine Peptidase 1; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D055213:Retinal Pigment Epithelium / Q000378:metabolism* / Q000473:pathology; D012697:Serine Endopeptidases / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000235:genetics
chemical_list: C525718:BMP2 protein, human; C525809:BMP4 protein, human; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D000074289:High-Temperature Requirement A Serine Peptidase 1; C474012:HTRA1 protein, human; D012697:Serine Endopeptidases