Effect of Wnt1 on the expression of Cyclin D1 protein in human corneal epithelial cells / 实用医学杂志

ABSTRACT

Objective To investigate the effect of Wnt1 on the expression of Cyclin D1 in human corneal epithelial cells and its related molecular mechanisms. Methods 12 T25 cell culture flasks were cultured after hu-man corneal epithelial cells anabiosis ,culture and continuous passage for 2 times. Culture flasks were divided into 3 groups with 4 culture flasks in each group. Twenty-five ng/mL and 50 ng/mL recombinant human Wnt1 protein were added in two of the groups,and one group without T-cell culture medium(Wnt1)was used as control. Cells cultured in T25 flask were taken from three groups at different time(6 h,24 h,48 h and 72 h). The total number of corneal epithelial cells in each group was calculated. Expression of Cyclin D1 in corneal epithelial cells was de-tected by Western blot. Results The expression of Cyclin D1 protein in the control group decreased gradually from 0 h to 48 h,and reached the lowest level at 48 h and increased at 72 h. Cyclin D1 protein expression in 25 ng/mL group at 6 h after Wnt1 was added was not detected,and Cyclin D1 protein expression in 50 ng/mL group in-creased. The expression of Cyclin D1 protein in 25 ng/mL group and 50 ng/mL group was significantly higher than that in control group at 24 h,48 h and 72 h,reaching the peak at 48 h and decreased at 72 h. Compared with the control group,the growth rate of corneal epithelial cells in 25ng/ml group and 50ng/ml group increased after Wnt1 was added. There was significant difference in 72 h,but no significant difference in 6h,24h and 48h. Conclu-sions The stimulation of Wnt1 protein can enhance the expression of Cyclin D1 in a certain time range,and has a positive correlation with Wnt1 protein. As one of the target genes of Wnt1 signaling pathway,Cyclin D1 may play an important role in the repair of corneal epithelial injury and its cell proliferation and differentiation.