June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
TGF-beta2 promotes RPE cell invasion into collagen gel by mediating urokinase-type plasminogen activator (uPA) expression
Author Affiliations & Notes
  • Koji Sugioka
    Ophthalmology, Kinki University Faculty of Medicine, Osakasayama, Japan
  • Aya Kodama
    Ophthalmology, Kinki University Faculty of Medicine, Osakasayama, Japan
  • Koji Yoshida
    Biochemistry, Kinki University Faculty of Medicine, Osakasayama, Japan
  • Kiyotaka Okada
    Physiology and Regenerative Medicine, Kinki University Faculty of Medicine, Osakasayama, Japan
  • Shunji Kusaka
    Ophthalmology, Sakai Hospital, Kinki University Faculty of Medicine, Sakai, Japan
  • Chota Matsumoto
    Ophthalmology, Kinki University Faculty of Medicine, Osakasayama, Japan
  • Yoshikazu Shimomura
    Ophthalmology, Kinki University Faculty of Medicine, Osakasayama, Japan
  • Footnotes
    Commercial Relationships Koji Sugioka, None; Aya Kodama, None; Koji Yoshida, None; Kiyotaka Okada, None; Shunji Kusaka, None; Chota Matsumoto, None; Yoshikazu Shimomura, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2012. doi:
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      Koji Sugioka, Aya Kodama, Koji Yoshida, Kiyotaka Okada, Shunji Kusaka, Chota Matsumoto, Yoshikazu Shimomura; TGF-beta2 promotes RPE cell invasion into collagen gel by mediating urokinase-type plasminogen activator (uPA) expression. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2012.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Transforming growth factor-beta (TGF-beta) is one of the main epithelial-mesenchymal transition (EMT)-inducing factors and TGF-beta-induced EMT promotes cell migration and invasion. In this study, we investigated the role of urokinase-type plasminogen activator (uPA) during TGF-beta-induced RPE cell invasiveness.

Methods: ARPE-19 cells were cultured and stimulated with TGF-beta2. uPA and uPA receptor (uPAR) expressions during TGF-beta-induced EMT process in ARPE-19 cells were evaluated by real-time RT-PCR and fibrin zymography. Effects of TGF-beta inhibitor (SB431542) on the expression of uPA and uPAR by TGF-beta2 stimulation were also evaluated. IAsys-binding assay was performed to investigate the binding ability of uPA to the surface of ARPE-19 cells. For invasion assay, ARPE-19 cells were seeded onto the Transwell chambers and allowed to invade into collagen matrix in the presence of TGF-beta2 alone or TGF-beta2+u-PA inhibitor.

Results: TGF-beta2 enhances expression of uPA and uPAR in ARPE-19 cells and secretion of uPA. SB431542 significantly suppressed TGF-beta2-stimulated uPA expression and secretion but not uPAR expression in ARPE-19 cells. ARPE-19 cells pretreated and non-pretreated with TGF-beta2 bound to uPA in a time-dependent or cell-number-dependent fashion. However, the binding ability of uPA to ARPE-19 cells was enhanced more greatly with ARPE-19 cells pretreated with TGF-beta2. TGF-beta2 treatment induced the invasiveness of ARPE-19 cells into collagen gel. TGF-beta2+u-PA inhibitor treatment strongly inhibited the invasiveness of ARPE-19 cells.

Conclusions: TGF-beta-induced RPE cell invasiveness occurs via its regulation of uPA. These results suggest that uPA mediates TGF-beta-induced RPE cell invasion in collagen gel matrix.

Keywords: 701 retinal pigment epithelium • 688 retina • 512 EMT (epithelial mesenchymal transition)  
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