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Inhibition of cell proliferation and migration after HTRA1 knockdown in retinal pigment epithelial cells

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Abstract

Purpose

The purpose of this study was to investigate the role of HtrA serine peptidase 1 (HTRA1) in the proliferation and migration of cells of the human retinal pigment epithelial cell line ARPE-19, and the possible mechanisms involved.

Methods

ARPE-19 cells were transduced by a recombinant lentiviral vector carrying HTRA1-shRNA to knockdown HTRA1 expression. Subsequent HTRA1 gene and HTRA1 protein levels in these cells and control cells were detected by quantitative real-time PCR and Western blot, respectively. Changes in cell proliferation and migration associated with the inhibition of HTRA1 expression were assessed, as well as changes in the mRNA levels of transforming growth factor beta 1 (TGFB1), bone morphogenetic protein 4 (BMP4), and bone morphogenetic protein 2 (BMP2).

Results

The recombinant lentivirus carrying HTRA1-shRNA was successfully generated, as evidenced by reduced levels of HTRA1 mRNA and HTRA1 protein in ARPE-19 cells. The knockdown of HTRA1 in ARPE-19 cells was associated with reduced cellular proliferation and migration, and increased mRNA levels of TGF-β1, BMP4, and BMP2.

Conclusions

Silence of the HTRA1 gene was associated with significantly higher levels of TGF-β1, BMP4, and BMP2 mRNA and reduction in the proliferation and migration of ARPE-19 cells.

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Acknowledgments

This research is supported by the National Natural Science Foundation of China Grant 81070734, the Beijing Education Commission Grant KZ201110025028, and the Beijing Municipal Health Bureau Grant 2009208.

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Authors and Affiliations

  1. Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology and Visual Science Key Laboratory, No. 1 Dongjiaominxiang, Dongcheng District, Beijing, China

    Xueting Pei, Kai Ma, Jun Xu, Ningli Wang & Ningpu Liu

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  1. Xueting Pei
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  2. Kai Ma
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  3. Jun Xu
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Correspondence to Xueting Pei.

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Pei, X., Ma, K., Xu, J. et al. Inhibition of cell proliferation and migration after HTRA1 knockdown in retinal pigment epithelial cells. Graefes Arch Clin Exp Ophthalmol 253, 565–572 (2015). https://doi.org/10.1007/s00417-014-2901-2

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  • DOI: https://doi.org/10.1007/s00417-014-2901-2

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