Influence of RGDRGD peptide on proliferation and adhesion of human lens epithelial cells in vitro / 中华实验眼科杂志

ABSTRACT

Background RGD is a small molecular multiple peptide containing Arg-Gly-Asp with an important role in inhibiting the adhesion,migration and neovascularization of tumor.Our previous study determined that RGD can suppress the adhesion and proliferation of lens epithelial cells(LECs),and RGDRGD may be of a stronger effect. Objective Present study was to investigate and compare the effect of RGDRGD peptide on the proliferation and adhesion of immortalized human LECs(HLEB-3)in vitro. Methods Human LECs harvested by trpsin-EDTA were suspended in DMEM medium with serial dilutions of RGD peptide and RGDRGD peptide(from 1000 mg/L to 250 mg/L)at 37℃ for 15 minutes as experimental group,and the HLECs cultured by common culture medium were used as the control group.The cells were then seeded into the 96-well plates with precoated fibmnectin (FN)and I collagen at the density of 2×104/ml.MTT stainingcolorimetry was used to measure the adhesion rates of lens epithelial cells cultured in different concentrations RGDRGD and RGD peptides after 1 hour.Cells were seeded into the 96-well plates for 24 hours at 37℃ in 5% CO2.Medium was then replaced with DMEM overnight.Subsequently,the cells were treated with serial dilutions of RGD and RGDRGD(from 2000 mg/L to 250 ms/L)dissolved in DMEM medium plus 20% fetal bovine serum.The inhibition of RGDRGD and RGD on the adhesion and proliferation of Human LECs was analyzed by MTT aher 24,48 and 72 hours. Results The inhibition rate of RGD peptide on the adhesion of LECs was gradually enhanced with the increase of concentration with the significant difference among the different concentrations groups(F=1089.56,P＜0.01),and the statistically significant elevation in inhibitory rate was found in RGDRGD peptide compared with RGD peptide(P＜0.01).The inhibition rate of RGDRGD peptide on the proliferation of LECs wag gradually increased with the increase of concentration with the significant difference among the different concentrations groups with a strongest effect in 1000 ms/L group(F=127.31,P＜0.01),and the much stronger inhibition Wag Been in RGDRGD peptide(F=1589.85,P＜0.01).The suppression rate of RGDRGD on LECs proliferation Wag much stronger with the prolong of time(F=1606.43,P＜0.01). Conclusion RGDRGD peptide and RGD peptide have inhibitory effect on adhesion and proliferation of human LECs in a dose-and time-dependent manner.Effect of RGDRGD peptide is much stronger than RGD peptide.These results imply that RGDRGD peptide and RGD peptide have the important role for prevention of PCO.