Recombined adenovirus of p21 inhibit the proliferation of Rhesus retinal vascular endothelial cells in vitro

Abstract

Objective To observe the inhibitory effect of recombined adenovirus of p21 (rAd-p21) on the proliferation of Rhesus retinal vascular endothelial cell line (RF/6A). MethodsRF/6A cells were cultured in vitro which divided into phosphate buffered solution (PBS) group, rAd-p21 transfection group and negative control (NC) group. Plasmid vectors were transfected into RF/6A cells. The expression of p21mRNA and protein in RF/6A cells were measured by RT-PCR and Western blot respectively. The cell cycle distribution was analyzed by flow cytometry. Endothelial-cell tube formation assay was performed in Matrigel. Results The expression of p21 mRNA and protein in rAd-p21 transfection group were higher than that in PBS and negative control group. The cell cycle distribution showed that the proportion of G0/G1 cells in rAd-p21 transfection group[(67.45 ± 11.61) ％]was apparently higher than that in the negative control group[(41.55±8.99)％]and PBS group[(40. 76±6.66) ％](F=21.284, P =0. 000 ). The number of endothelial-cell tubes in the rAd-p21 transfection group (3.86±1.21) was apparently less than that in the negative control group (7.62±2.69) and PBS group (8.25±3.19) (F=7.138,P=0.004).Conclusions The p21 mRNA and protein can stably express in RF/6A cells after rAd-p21 transfection.RAd-p21 can inhibit the proliferation of RF/6A cells. Key words: Retinal neovascularization/ prevention & control; Gene transfer techniques; Transfection; Animal experimentation