Qiu-Li Zhang1, Wei Wang2, Jin Li3, Shi-Yuan Tian4, Tian-Zi Zhang5. 1. Department of Ophthalmology, Affiliated Hospital of Inner Mongolia University for the Nationalities, No. 1742, HuoLinHe Street, Tongliao, Neimenggu 028050, China. 2. Department of Ophthalmology, Tongliao City Hospital, No. 668, KeErQin Street, Tongliao, Neimenggu 028000, China. 3. Department of Infusion Center, The Children's Hospital of Changchun City, No.1321, Beian Road Kuancheng District, Changchun, Jilin 130000, China. 4. Department of Ophthalmology, Affiliated Hospital of Inner Mongolia University for the Nationalities, No. 1742, HuoLinHe Street, Tongliao, Neimenggu 028050, China. Electronic address: tiansyuan@163.com. 5. Department of Ophthalmology, Affiliated Hospital of Inner Mongolia University for the Nationalities, No. 1742, HuoLinHe Street, Tongliao, Neimenggu 028050, China. Electronic address: z_tianzi@163.com.
Abstract
BACKGROUND: Retinal ganglion cells (RGCs) are commonly experienced optic nerve diseases including glaucoma-induced injury that results in decrease of cell survival. However, the underlying mechanism remains to be elaborated. This present study was to focus on the miR-187 and Transforming growth factor-β (TGF-β) signal and investigated their roles in RGCs apoptosis and proliferation. METHODS: RGC-5 retinal ganglion cell line was chose in present study and subjected to miR-187 mimic or inhibitor transfection. Cell apoptosis was evaluated using flow cytometry-based Annexin V-PI assay. Cell proliferation was examined using CCK-8. Protein levels of Smad2/3/7 were determined using western blotting. RESULTS: miR-187 negatively regulated cell survival via inhibiting cell apoptosis and promoting cell proliferation. We observed that alteration expression of miR-187 is closely related to phosphorylation levels of Smad2 and Smad3. This correlation is associated with down-regulation of Smad7 induced by miR-187 via targeting Smad7 3'-UTR. From result of co-transfection of Smad7-plasmid and miR-187 mimic or siSmad7 and miR-187 inhibitor, we concluded that cell proliferation and apoptosis was mediated by miR-187/Smad7 axis. CONCLUSION: In summary, cell internal signal transduction, miR-187 regulating Smad7 expression, plays a vital role in retinal ganglion cell survival.
BACKGROUND: Retinal ganglion cells (RGCs) are commonly experienced optic nerve diseases including glaucoma-induced injury that results in decrease of cell survival. However, the underlying mechanism remains to be elaborated. This present study was to focus on the miR-187 and Transforming growth factor-β (TGF-β) signal and investigated their roles in RGCs apoptosis and proliferation. METHODS: RGC-5 retinal ganglion cell line was chose in present study and subjected to miR-187 mimic or inhibitor transfection. Cell apoptosis was evaluated using flow cytometry-based Annexin V-PI assay. Cell proliferation was examined using CCK-8. Protein levels of Smad2/3/7 were determined using western blotting. RESULTS:miR-187 negatively regulated cell survival via inhibiting cell apoptosis and promoting cell proliferation. We observed that alteration expression of miR-187 is closely related to phosphorylation levels of Smad2 and Smad3. This correlation is associated with down-regulation of Smad7 induced by miR-187 via targeting Smad7 3'-UTR. From result of co-transfection of Smad7-plasmid and miR-187 mimic or siSmad7 and miR-187 inhibitor, we concluded that cell proliferation and apoptosis was mediated by miR-187/Smad7 axis. CONCLUSION: In summary, cell internal signal transduction, miR-187 regulating Smad7 expression, plays a vital role in retinal ganglion cell survival.
Authors: Mohsen Ghanbari; Adriana I Iglesias; Henriët Springelkamp; Cornelia M van Duijn; M Arfan Ikram; Abbas Dehghan; Stefan J Erkeland; Caroline C W Klaver; Magda A Meester-Smoor Journal: Invest Ophthalmol Vis Sci Date: 2017-10-01 Impact factor: 4.799