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Effects of AFP-172 on COX-2-induced angiogenic activities on human umbilical vein endothelial cells

  • Basic Science
  • Published:
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Abstract

Purpose

To investigate the angiogenic effect of the free acid of tafluprost (AFP-172) on human umbilical vascular endothelial cells (HUVECs).

Methods

HUVECs cultured in the presence or absence of FP receptor antagonist (10 nM AL-8810) were exposed to escalating concentrations of 10−7, 10−6, 10−5, 10−4 and 10−3 M AFP-172 (the free acid of tafluprost). For cell proliferation assays, the numbers of cells were derived from a CellTiter96® Aqueous One Solution Cell Proliferation Assay (Promega) by Microplate reader (Bio-Rad, Benchmark). Endothelial cell migration was evaluated by a BD Biocoat™ Angiogenesis System using FluoroBlok ™ 24-well inserts (BD Biosciences, Bedford, MA). BioTek FLx800 fluorescence plate reader was used for quantitative measurement of fluorescently-labeled invasive vascular endothelial cells. Endothelial capillary-like tube formation was evaluated by BD Biocoat Angiogenesis System using Matrigel Matrix 96-well plate. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assess the gene expression of vascular endothelial growth factor (VEGF), cyclooxygenase-2 (COX-2) and endothelial nitric oxide synthase (eNOS). COX-2 protein was detected by immunofluorescent staining and Western blot assay. Student's t-test was used for statistical analysis.

Results

10−4 M AFP-172 treated cells stimulated the proliferation, migration and tube formation of HUVECs as compared to 10−5, 10−6, 10−7 M AFP-172 treated cells and control (P < 0.01). RT-PCR showed that incubation of HUVECs with 10−4 M AFP-172 stimulated the expression of COX-2 mRNA (P < 0.05). Western blot assay revealed that AFP-172 caused cells to increase in COX-2 protein at the concentrations of 10−4 M.

Conclusions

>AFP-172 showed the angiogenic effects on HUVECs at the concentrations of 10−4 M by inducing COX-2 protein.

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Acknowledgments

The authors wish to acknowledge the financial support of the Catholic Medical Center Research Foundation made in the program year of 2010.

Conflict of interest

No author has financial or proprietary interests in any material or method mentioned.

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Authors and Affiliations

  1. Department of Ophthalmology and Visual Science, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea

    Young Jung Roh, Young Gun Park, Seungbum Kang & Jung Il Moon

  2. Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea

    Soo Young Kim

  3. Department of Ophthalmology, Yeouido St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, #62 Yeouido-dong, Yeongdeungpo-gu, Seoul, 150-713, Korea

    Jung Il Moon

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  1. Young Jung Roh
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Correspondence to Jung Il Moon.

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Roh, Y.J., Park, Y.G., Kang, S. et al. Effects of AFP-172 on COX-2-induced angiogenic activities on human umbilical vein endothelial cells. Graefes Arch Clin Exp Ophthalmol 250, 1765–1775 (2012). https://doi.org/10.1007/s00417-012-2125-2

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  • DOI: https://doi.org/10.1007/s00417-012-2125-2

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