September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Expression of A Disintegrin and Metalloproteinase 17 (ADAM 17) in RPE cell culture is associated with apoptosis
Author Affiliations & Notes
  • Chen Liang
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, Sichuan, China
    Ophthalmology and Visual Sciences, University of British Columbia, Vancouver, British Columbia, Canada
  • Jing Z Cui
    Ophthalmology and Visual Sciences, University of British Columbia, Vancouver, British Columbia, Canada
  • Luba Kojic
    Brain Research Centre, University of British Columbia, Vancouver, British Columbia, Canada
  • William Jia
    Brain Research Centre, University of British Columbia, Vancouver, British Columbia, Canada
    Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada
  • Joanne A Matsubara
    Ophthalmology and Visual Sciences, University of British Columbia, Vancouver, British Columbia, Canada
  • Footnotes
    Commercial Relationships   Chen Liang, None; Jing Cui, None; Luba Kojic, None; William Jia, None; Joanne Matsubara, None
  • Footnotes
    Support  CIHR MOP 126195
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 6557. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Chen Liang, Jing Z Cui, Luba Kojic, William Jia, Joanne A Matsubara; Expression of A Disintegrin and Metalloproteinase 17 (ADAM 17) in RPE cell culture is associated with apoptosis. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6557.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : ADAM17 has been reported to be an indispensible regulator of almost every cellular event from proliferation to death. However, the role of ADAM17 in eye diseases is unknown. The purpose of this study is to investigate the expression and the function of ADAM17 in RPE cells in vitro and in postmortem eyes of AMD and non-AMD donors.

Methods : ARPE19 cells were stimulated with IL-1β, H2O2 or TNF-α to trigger ADAM17 activation for 24 hrs. Cells were then lysed, and samples extracted for total, cytoplasmic or nuclear protein. ADAM17 protein expression was assessed by western blot. Nuclear localization of ADAM17 was further evaluated by immunocytochemistry on culture slides. Apoptosis in stimulated ARPE19 cells was studied using a pan-caspase inhibitor, flow cytometry and annexin V/PI labeling. Results were analyzed by one-way ANOVA, p < 0.05.

Results : Total protein analysis revealed three forms of ADAM17: proform (130kD), mature form (100kD) and cytoplasmic tail form (40kD). Significant fold increase of the mature form was observed for IL-1β (1.8±0.45), H2O2 (1.5±0.04), TNF-α (2.0±0.28) stimulation; no significant changes were observed in the cytoplasmic tail form in any of the stimulation regimes. However, the cytoplasmic tail form was significantly increased in the nuclear protein material in all treatment groups (1.9 -4.0 fold). Cell fluorescence staining further confirmed ADAM17 signal in RPE nuclei. Nuclear cytoplasmic tail form was reduced with pretreatment of pan-caspase inhibitor, which blocks caspase-dependent apoptosis. Corroborative immunohistochemistry data were observed from sections of human AMD eyes, in which increased ADAM17 levels were found in the AMD eyes (N=13) and in the nuclear compartment when compared to age-matched controls (N=7).

Conclusions : This is the first study to demonstrate a nuclear localization of ADAM17 in RPE cell under stress in vitro and its role in RPE apoptosis. Additionally, the presence of ADAM17 in RPE nuclei of postmortem eyes is suggestive of a possible association with AMD disease processes.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×