Tissue microarray analysis of cyclin-dependent kinase inhibitors p21 and p16 in Fuchs dystrophy.
Tissue microarray analysis of cyclin-dependent kinase inhibitors p21 and p16 in Fuchs dystrophy.
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DOI：
10.1097/ico.0b013e31826f324e
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发表时间：
2013-04
期刊：
影响因子：
2.8
通讯作者：
Jun AS
中科院分区：
文献类型：
--
作者：
Matthaei M;Lackner EM;Meng H;Hicks JL;Meeker AK;Eberhart CG;Jun AS
研究组织微阵列（TMA）技术在角膜疾病中的新应用，并报道衰老相关的细胞周期蛋白依赖性激酶抑制剂p21和p16在Fuchs内皮性角膜营养不良（FECD）中的蛋白表达改变。从50例FECD患者和5例角膜圆锥（KC）患者穿透性角膜移植术后的角膜扣、10例非病理角膜解剖球和非眼对照标本的石蜡包埋组织中生成包括208个核的TMA。TMA切片免疫标记p21和p16，并使用9级评分系统（0-8）进行分析。通过对单个组织切片进行免疫标记来验证结果。与对照组相比，FECD标本中角膜内皮p21和p16的表达水平是主要的结局指标。TMA免疫组化分析显示feecd标本中细胞核p21表达水平升高（p<0.05），内皮细胞p16表达模式改变。全组织切片免疫标记显示两种蛋白的内皮过表达有统计学意义（p21和p16， p<0.05）。本研究介绍了TMA技术作为角膜组织分子高通量分析的有价值的工具。p21和p16在遗传未分化FECD患者的角膜内皮中过表达，支持细胞衰老在FECD发病机制中的作用。
To investigate the novel application of tissue microarray (TMA) technology to corneal disease and to report altered protein expression of senescence-associated cyclin-dependent kinase inhibitors p21 and p16 in Fuchs endothelial corneal dystrophy (FECD). A TMA including 208 cores was generated from paraffin-embedded tissues including corneal buttons of 50 FECD and 5 keratoconus (KC) patients retrieved post penetrating keratoplasty, 10 autopsy globes with non-pathologic corneas, and non-ocular control specimens. TMA sections were immunolabeled for p21 and p16 and analyzed using a nine-grade scoring system (0–8). Result validation was performed by immunolabeling of individual whole tissue sections. Corneal endothelial p21 and p16 expression levels in FECD specimens compared to controls served as main outcome measures. TMA immunohistochemical analysis disclosed increased endothelial expression levels of nuclear p21 in FECD specimens (p<0.05) and an altered endothelial p16 expression pattern. Immunolabeling of whole tissue sections showed statistically significant endothelial overexpres-sion of both proteins (p21 and p16, p<0.05). The present study introduces TMA technology as a valuable tool for molecular high-throughput profiling of corneal tissues. It demonstrates p21 and p16 overexpression in the corneal endothelium of genetically undifferentiated FECD patients supporting a role of cellular senescence in the pathogenesis of FECD.