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Contribution of the clock gene DEC2 to VEGF mRNA upregulation by modulation of HIF1α protein levels in hypoxic MIO-M1 cells, a human cell line of retinal glial (Müller) cells

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Abstract

Purpose

Clock genes are components of the molecular clock. Their malfunction is thought to increase the risk of numerous diseases, including cancer. Vascular endothelial growth factor (VEGF) has a pivotal role in angiogenesis, and its expression levels are controlled by clock genes in tumor cells. Ophthalmic diseases such as age-related macular degeneration, proliferative diabetic retinopathy, and neovascular glaucoma are also associated with abnormal angiogenesis followed by upregulation of VEGF in the eye. In the present study, we aimed to uncover the relationship between clock genes and VEGF in the eye.

Study design

Laboratory investigation

Methods

Oxygen-induced retinopathy (OIR) mice were prepared to mimic hypoxic conditions in the eye. Deferoxamine (DFO) was used to mimic hypoxic conditions in human Müller cell line MIO-M1 cells. Expression levels of mRNA and protein were quantified by quantitative reverse transcription polymerase chain reaction and Western blot analysis, respectively.

Results

In the retinas of OIR mice, the expression levels of Vegf and the clock gene Dec2 increased transiently, and their temporal profiles were correlated. Knockdown of DEC2 resulted in a significant (26.7%) reduction of VEGF expression in MIO-M1 cells under hypoxia-mimicking conditions induced by DFO (P < .05). Levels of HIF1α protein were also reduced significantly, by 60.2%, in MIO-M1 cells treated with siRNA against the DEC2 gene (P < .05). Moreover, HIF1α levels showed a significant (2.5-fold) increase in MIO-M1 cells overexpressing DEC2 (P < .05).

Conclusion

DEC2 could upregulate retinal VEGF gene expression through modulation of HIF1α levels under hypoxic conditions.

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Acknowledgements

This work was supported by a Grant-in-Aid for Young Scientists (B) (no. 26861456) from the Japan Society for the Promotion of Science.

Author information

Authors and Affiliations

  1. Department of Ophthalmology, Faculty of Medicine, Oita University, 1-1 Yufu-shi, Oita, 879-5593, Japan

    Naoki Kusunose, Takahiro Akamine, Kenichi Kimoto & Toshiaki Kubota

  2. Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan

    Naoki Kusunose, Takahiro Akamine, Sai Yasukochi & Shigehiro Ohdo

  3. Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    Yoshiyuki Kobayashi & Shigeo Yoshida

  4. Department of Glocal Healthcare, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan

    Naoya Matsunaga & Satoru Koyanagi

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  1. Naoki Kusunose
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  2. Takahiro Akamine
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Corresponding author

Correspondence to Naoki Kusunose.

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Conflicts of interest

N. Kusunose, None; T. Akamine, None; Y. Kobayashi, None; S. Yoshida, None; K. Kimoto, None; S. Yasukochi, None; N. Matsunaga, None; S. Koyanagi, None; S. Ohdo, None; T. Kubota, None.

Additional information

Corresponding author: Naoki Kusunose

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Kusunose, N., Akamine, T., Kobayashi, Y. et al. Contribution of the clock gene DEC2 to VEGF mRNA upregulation by modulation of HIF1α protein levels in hypoxic MIO-M1 cells, a human cell line of retinal glial (Müller) cells. Jpn J Ophthalmol 62, 677–685 (2018). https://doi.org/10.1007/s10384-018-0622-5

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  • DOI: https://doi.org/10.1007/s10384-018-0622-5

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