June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Transcription factor 4 Regulates Mitochondrial Function and Energy Metabolism in cultured Human Corneal Endothelial Cells
Author Affiliations & Notes
  • Young Joo Shin
    Ophthalmology, Hallym University College of Medicine, Hallym Medical Center, Seoul, Korea (the Republic of)
  • Tae-Young Chung
    Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea (the Republic of)
  • Soonil Kwon
    Ophthalmology, Hallym University College of Medicine, Hallym Medical Center, Seoul, Korea (the Republic of)
  • Joon-Young Hyon
    Seoul National University Bundang Hospital, Seongnam, Korea (the Republic of)
  • Footnotes
    Commercial Relationships   Young Joo Shin, None; Tae-Young Chung, None; Soonil Kwon, None; Joon-Young Hyon, None
  • Footnotes
    Support  This research was supported by the National Research Foundation (NRF) grant funded by the Korea government (NRF-2015R1D1A1A09058505) and by Hallym University Research Fund 2016 (HURF-2016-12).
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1440. doi:
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    • Get Citation

      Young Joo Shin, Tae-Young Chung, Soonil Kwon, Joon-Young Hyon; Transcription factor 4 Regulates Mitochondrial Function and Energy Metabolism in cultured Human Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1440.

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

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Abstract

Purpose : To investigate the role of TCF4 in the mitochondrial functions of human corneal endothelial cells (HCECs).

Methods : HCECs were cultured, and TCF4 was blocked by siRNA (si-TCF4) or activated using CRISPR/dCas9 systems for TCF4 (pl-TCF4). Cell viability was assessed using a WST-8 assay kit, and mitochondrial membrane potential (ΔΨm) was measured using the JC-1 dye. Intracellular reactive oxygen species (ROS) formation was measured using a DCF-DA probe. Protein expression was evaluated by western blotting. Induction of autophagy and apoptosis were also evaluated.

Results : A mitochondrial viability stain demonstrated that mitochondrial viability was reduced by blocking TCF4 and elevated by its overexpression (p <0.05). ΔΨm was measured using JC-1, and was decreased by blocking TCF4 and elevated by TCF4 overexpression (p <0.05). Intracellular ROS production was evaluated by dichlorofluorescein (DCF) fluorescence. Intracellular ROS formation increased with TCF4 inhibition and decreased with its overexpression (p < 0.05). Relative adenosine triphosphate (ATP) production decreased upon TCF4 inhibition, but was elevated by its overexpression (p = 0.009 and 0.026). Activation of AMP-activated protein kinase (AMPK) decreased with inhibition of TCF4 expression, however TCF4 overexpression has the opposite effect (p < 0.05). In contrast, sirtuin 1 (SIRT1) expression was elevated by blocking TCF4 and reduced by its overexpression (p <0.05).

Conclusions : TCF4 regulates mitochondrial function and energy production in cultured HCECs.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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