Li Jing, Riau Andri K, Setiawan Melina, Mehta Jodhbir S, Ti Seng-Ei, Tong Louis, Tan Donald T H, Beuerman Roger W
Department of Ophthalmology, Xinhua Hospital, Shanghai Jiao Tong University, China.
Mol Vis. 2011;17:2263-71. Epub 2011 Aug 20.
To study the expression and cellular distribution of multiple S100A genes and proteins in normal corneal-limbal epithelium and ocular surface squamous cell carcinoma (SCC) tissue.
Normal corneal-limbal tissue was obtained from the Lions Eye Bank, Tampa, FL. Ocular surface SCC tissues were excised from patients undergoing surgery at Singapore National Eye Centre. S100A mRNA expression was measured by quantitative PCR. S100 protein distribution was determined by immunofluorescent staining analysis.
Twelve S100 mRNAs were identified in human corneal and limbal epithelial cells. S100A2, A6, A8, A9, A10, and A11 mRNA was expressed at high level, while S100A1, A3, A4, A5, A6, A7, and A12 mRNA expression was low. The intracellular localization of S100A2, A6, A8, A9, A10 and A11 protein was determined in normal corneal-limbal and SCC tissues. S100A2 and S100A10 proteins were enriched in basal limbal epithelial cells of the normal tissue. S100A8 and S100A9 were found only at the surface of peripheral corneal and limbal epithelium. S100A6 was uniformly found at the plasma membrane of corneal and limbal epithelial cells. S100A11 was found at the supralayer limbal epithelial cells adjacent to the conjunctiva. SCC tissue showed typical pathological changes with expression of cytokeartin (CK) 14 and CK4 in the epithelial cells. All SCC epithelial cells were positive of S100A2, S100A10, S100A6 and S100A11 staining. Intracellular staining of S100A8 and S100A9 was found in several layers of SCC epithelium. Expression of S100A2 and S100A10 decreased dramatically in cultured limbal epithelial cells with increased passaging, which was accompanied by a small increase of S100A9 mRNA, with no changes of S100A8 gene expression. Serum and growth hormone depletion of the culture serum caused a small reduction of S100A2 and S100A10 gene expression, which was accompanied by a small increase of S100A9 mRNA while no changes of S100A8 expression was measured.
Normal corneal and limbal epithelial cells express a broad spectrum of S100 genes and proteins. Ocular surface SCC express high levels of S100A2, S100A10, S100A8 and S100A9 proteins. The expression of S100A2 and S100A10 is associated with limbal epithelial cell proliferation and differentiation.
研究多种S100A基因和蛋白在正常角膜缘上皮及眼表鳞状细胞癌(SCC)组织中的表达及细胞分布。
正常角膜缘组织取自佛罗里达州坦帕市的狮子眼库。眼表SCC组织取自新加坡国立眼科中心接受手术的患者。通过定量PCR检测S100A mRNA表达。通过免疫荧光染色分析确定S100蛋白分布。
在人角膜和角膜缘上皮细胞中鉴定出12种S100 mRNA。S100A2、A6、A8、A9、A10和A11 mRNA高表达,而S100A1、A3、A4、A5、A6、A7和A12 mRNA低表达。在正常角膜缘和SCC组织中确定了S100A2、A6、A8、A9、A10和A11蛋白的细胞内定位。S100A2和S100A10蛋白在正常组织的角膜缘基底上皮细胞中富集。S100A8和S100A9仅在外周角膜和角膜缘上皮表面发现。S100A6均匀分布于角膜和角膜缘上皮细胞的质膜。S100A11在结膜相邻的角膜缘上皮上层细胞中发现。SCC组织显示典型病理变化,上皮细胞中细胞角蛋白(CK)14和CK4表达。所有SCC上皮细胞S100A2、S100A10、S100A6和S100A11染色均为阳性。在SCC上皮的几层细胞中发现S100A8和S100A9的细胞内染色。随着传代次数增加,培养的角膜缘上皮细胞中S100A2和S100A10表达显著降低,同时S100A9 mRNA略有增加,S100A8基因表达无变化。培养血清中血清和生长激素的缺乏导致S100A2和S100A10基因表达略有降低,同时S100A9 mRNA略有增加,而未检测到S100A8表达变化。
正常角膜和角膜缘上皮细胞表达多种S100基因和蛋白。眼表SCC高表达S100A2、S100A10、S100A8和S100A9蛋白。S100A2和S100A10的表达与角膜缘上皮细胞增殖和分化相关。