Wnt1蛋白对人角膜上皮细胞Cyclin D1蛋白表达的影响及意义  

Effect of Wnt1 on the expression of Cyclin D1 protein in human corneal epithelial cells

作　　者：陈梨萍[1] 李科[1] 郑振优[1] 宋绪华[1] 李雷[1] 赵楠楠[1] 

机构地区：[1]海南医学院第一附属医院眼科,海口市570102

出　　处：《实用医学杂志》2017年第11期1731-1734,共4页The Journal of Practical Medicine

基　　金：海南省医药卫生科技项目基金资助(编号:1341320;37A2010);海南医学院第一附属医院青年培育项目基金资助(编号:HYFYPY201603)

摘　　要：目的:探讨Wnt1蛋白对人角膜上皮细胞Cyclin D1蛋白表达的影响及其相关分子机制。方法:人角膜上皮细胞经复苏、培养,连续传代2次后共培养12个T25细胞培养瓶,4个培养瓶一组,共3组,分别加入25 ng/m L的重组人Wnt1蛋白和50 ng/m L的重组人Wnt1蛋白,一组不加入重组人Wnt1蛋白作为对照,于不同时间点(6、24、48和72 h)各取一个T25培养瓶的细胞,计算各组角膜上皮细胞总数并采用Western blot方法检测角膜上皮细胞中Cyclin D1蛋白的表达情况。结果:对照组Cyclin D1蛋白的表达在0~48 h内随着时间的延长呈现逐步降低的趋势,在48 h达到最低状态,在72 h有所提高。加入Wnt1后6 h,25 ng/m L组未检测到Cyclin D1蛋白表达,50 ng/m L组Cyclin D1蛋白表达有所提高。在24,48和72 h时间点,25 ng/m L组和50 ng/m L组Cyclin D1蛋白表达均提高,48 h达到最高峰,72 h有所回落。添加Wnt1后与对照组比较,25 ng/m L组及50 ng/m L组角膜上皮细胞生长速度均加快,但在6、24及48 h时间点差异无显著性,在72 h差异有显著性。结论:Wnt1蛋白的刺激可在一定时间范围内增强人角膜上皮细胞Cy-clin D1的表达,且与Wnt1蛋白量呈正相关。Cyclin D1的表达强弱,在角膜上皮损伤修复及其细胞增殖分化过程中可能起重要作用。Objective To investigate the effect of Wntl on the expression of Cyclin D1 in human corneal epithelial ceils and its related molecular mechanisms. Methods 12 T25 cell culture flasks were cultured after human 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 Wntl protein were added in two of the groups, and one group without T-cell culture medium （Wntl） 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 D 1 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 Wntl 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 Wntl was added. There was significant difference in 72 h, but no significant difference in 6h, 24h and 48h. Conclu- sions The stimulation of Wnt 1 protein can enhance the expression of Cyclin D 1 in a certain time range, and has a positive correlation with Wntl protein. As one of the target genes of Wntl signaling pathway, Cyclin D1 may play an important role in the repair of corneal epithelial injury and its cell proliferation and differentiation.

关 键 词：CYCLIN D1 人角膜上皮细胞 角膜 Wntl 

分 类 号：R772.2[医药卫生—眼科]