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X. Ding, L. Lei, T. Li, S. Xie, H. Ma, B. Li, S. Tang; Norrin Induced the Proliferation and Migration of Cultured Human Retinal Capillary Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):3431.
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To isolate, cultivate and identify human retinal capillary endothelial cells (HRCECs), and to assess the effects of high expression of Norrin gene on the proliferation, cell cycle, migration and apoptosis of HRCECs.
The cultured cells were identified with anti-factor VIII related antigen and W-P body by transmission electron microscope(TEM). AP-3myc-hNorrin/pRK5 were transfected into cultured HRCECs in vitro by lipofectamine2000. Their effects on mRNA of Norrin and protein expression of myc tag were measured by RT-PCR , Immunohistochemistry and Western blot,respectively. Its effects on the cell proliferation, cell cycle, migration and apoptosis were detected. The effects on cell migration were evaluated by TRANSWELL .
In comparison with those of the controls , the Norrin expression in transferred cells was significantly increased on mRNA and protein level(showed by the myc tag) after 48 hours. The cell number of transfered group was larger than that of the control group with statistical significant differences(P value 0.001), Flow cytometry showed the cells in G2/M phase were mainly increased (P value 0.001), and the cell migration was significantly improved in HRCECs transfected with Norrin gene(P value 0.001). The early and late stage apoptotic percentage of HRCECs in transferred group was significantly lower than that of the control group(P value 0.001).
Norrin gene can improve the proliferation and migration of HRCECs, and drive the cell cycle from G0 phase to G2 phase.,At the same time, Norrin gene can inhibit the apoptosis of HRCECs, Norrin may have an important role in the retinal angiogenesis ,which may provide a new gene target in the treatment of retinal vascular disorders.
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