Effects of the hypoxia-induced human retinal pigment epithelial cells which transfected by adenovirus-mediated Slit2 and adenovirus-mediated Slit2 ShRNA on the proliferation of human choroidal microvascular endothelial cells

Abstract:

Objective：To explore the effect of the hypoxia-induced human retinal pigment epithelial cells（RPE） which transfected by adenovirus-mediated Slit2 and adenovirus-mediated Slit2 ShRNA on the proliferation of human choroidal microvascular endothelial cells（HCMEC），to explore the possible role of Slit2 in choroidal neovascularization（CNV） and to provide new ideas for the treat－ment of choroidal neovascularization. Methods：RPE cells and HCMEC cells were identified and cultured in vitro. Hypoxic conditions were achieved by adding 200 ?滋mol/l cobalt chloride to medium to mimic hypoxia. RPE cells and HCMEC cells were cultured in a contact co-culture system by Transwell chamber. The hypoxia RPE cells were randomly divided into Slit2 treated group（adding Slit2RNA），Slit2 shRNA treated group（adding Slit2 shRNA），empty adenovirus group（adding empty adenovirus），hypoxia group. CCK8 assay was carried out to determine the proliferation of HCMEC cells after 12，24，48 h. Results：There were statistically signifi－cant differences among different groups（F=98.122，P=0.000） and different time points（F=3388.913，P=0.000）. The interaction be－tween different groups and time points were also different（F=82.863，P=0.000）. The absorbance（A） value was higher in Slit2 treated group than in other groups（hypoxia group：P=0.001；other groups：P=0.000）. The A value was lower in Slit2 shRNA treated group than in other groups at 24 h and 48 h（at 48 h：hypoxia group，P=0.003；empty adenovirus group，P=0.008；other groups，P=0.000）. Conclusion：High-expression of Slit2 can significantly promote the proliferation of HCMEC cells. Silencing Slit2 shRNA expression in RPE cell can significantly inhibit the proliferation of HCMEC cells.