Wnt1蛋白对人角膜上皮细胞Cyclin D1蛋白表达的影响及意义

发布时间：2018-06-07 06:33

本文选题：Cyclin + D　；参考：《实用医学杂志》2017年11期

【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。
[Abstract]:Aim: to investigate the effect of Wnt1 protein on the expression of Cyclin D1 protein in human corneal epithelial cells and its molecular mechanism. Methods: human corneal epithelial cells were resuscitated, cultured, subcultured for 2 times and then cultured in 12 T25 cell culture flasks, 4 flasks and 3 groups, respectively, with 25 ng/m L recombinant human Wnt1 protein and 50 ng/m L recombinant human Wnt1 protein. One group did not add recombinant human Wnt1 protein as control group, one cell in T25 culture bottle was taken at different time points, and the total number of corneal epithelial cells in each group was calculated, and the expression of Cyclin D1 protein in corneal epithelial cells was detected by Western blot method. Results: the expression of Cyclin D1 protein in the control group decreased gradually with the prolongation of time within 48 h, reached the lowest level at 48 h, and increased at 72 h. The expression of Cyclin D1 protein was not detected in 25 ng/m L group 6 h after adding Wnt1. The expression of Cyclin D1 protein was increased in 50 ng/m L group. The expression of Cyclin D1 protein in 25 ng/m L group and 50 ng/m L group increased at 24 h, 48 h and 72 h, respectively. The expression of Cyclin D1 protein increased at 48 h and reached the peak at 48 h and decreased at 72 h. Compared with the control group, the growth rate of corneal epithelial cells in 25 ng/m L group and 50 ng/m L group was faster than that in the control group, but there was no significant difference at 6h 24 and 48 h, but there was significant difference at 72 h. Conclusion the expression of Cy-clin D1 in human corneal epithelial cells could be enhanced by the stimulation of WNT1 protein, and the expression of Wnt1 protein was positively correlated with the expression of cyclin D1. It may play an important role in corneal epithelial injury repair and cell proliferation and differentiation.
【作者单位】：海南医学院第一附属医院眼科;
【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)
【分类号】：R772.2

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