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目的 检测体外培养角膜上皮细胞Ｐａｘ－６基因的达，探讨维持角膜缘干细胞特性的基因因素。 方法 地高辛标记ｃＤＮＡ探针进行ＤＮＡ印迹杂交和蛋白质免疫印迹，检测培养的角膜上皮细胞中Ｐａｘ－６基因表达状况。 结果 ＥｃｏＲ Ⅰ酶切角膜缘上皮细胞中９．１ｋｂｐ的ＤＮＡ片断与探针序列相同，其中原位和原代培养细胞、传第１代明显测出，角膜中央上皮细胞的原位和原代细胞中也能测出。Ｗｅｓｔｅｒｎ Ｂｌｏｔ发现抗血清１１认识相对分子质量为８１×１０３和５５×１０３两种角膜上皮细胞蛋白，抗血清１３认识６０×１０３的角膜上皮细胞蛋白，原位和原代、传第１代角膜缘干细胞和角膜中央上皮细胞均能够测出，角膜缘传第３代细胞和角膜基质细胞不表达。 结论 角膜缘干细胞和角膜中央上皮细胞均具有Ｐａｘ－６基因，该基因的正常表达是维持角膜上皮细胞特性的必要条件。
Objective To detect the expression of Pax-6 gene in cultured corneal epithelial cells in vitro and to explore the genetic factors that maintain the characteristics of limbal stem cells. Methods Digoxigenin-labeled cDNA probes were used to detect the Pax-6 gene expression in cultured corneal epithelial cells by Southern blotting and Western blotting. Results The DNA fragment of 9.1kbp in the corneal limbal epithelial cells was the same as that of EcoR Ⅰ . The primary and primary cultured cells passed the first passage and were obviously detected in situ and primary Cells can be measured. Western Blot found that the antisera 11 recognized two kinds of corneal epithelial cell proteins with relative molecular masses of 81 × 103 and 55 × 103, antisera 13 recognized 60 × 103 corneal epithelial cell protein, in situ and primary, passing on the first generation limbal margin Stem cells and central corneal epithelial cells were able to be detected, the third generation of corneal limbal cells and corneal stromal cells do not express. Conclusion Both limbal stem cells and central corneal epithelial cells have Pax-6 gene. The normal expression of this gene is a necessary condition to maintain the characteristics of corneal epithelial cells.