Sotozono C, Kinoshita S, Kita M, Imanishi J
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Japan.
Jpn J Ophthalmol. 1994;38(4):353-9.
The expression of keratinocyte growth factor (KGF) messenger RNA (mRNA) was examined in both ex vivo and in vitro specimens of human skin, conjunctiva and cornea. Total cellular mRNA was extracted from human skin, conjunctiva, cornea, cultured skin fibroblasts, cultured conjunctival fibroblasts and cultured keratocytes. Oligo dT-primed complementary DNA (cDNA) was synthesized using each RNA sample as a template. Polymerase chain reaction (PCR) was used to amplify the DNA sequence of KGF using each cDNA sample as a template. PCR products were then digested with restriction enzymes in order to determine the KGF prototype. In all samples, PCR products of the expected size for KGF were detected. The resulting restriction pattern proved that these products were identical to KGF prototype I, ie, authentic KGF. In conclusion, human keratocytes and conjunctival fibroblasts express mRNA coding for KGF prototype I, ie, authentic KGF, in both ex vivo and in vitro specimens.
在人皮肤、结膜和角膜的离体和体外标本中检测了角质形成细胞生长因子（KGF）信使核糖核酸（mRNA）的表达。从人皮肤、结膜、角膜、培养的皮肤成纤维细胞、培养的结膜成纤维细胞和培养的角膜细胞中提取总细胞mRNA。以每个RNA样本为模板合成寡聚dT引发的互补DNA（cDNA）。以每个cDNA样本为模板，使用聚合酶链反应（PCR）扩增KGF的DNA序列。然后用限制性内切酶消化PCR产物，以确定KGF原型。在所有样本中，均检测到KGF预期大小的PCR产物。所得的限制性图谱证明这些产物与KGF原型I相同，即真正的KGF。总之，人角膜细胞和结膜成纤维细胞在离体和体外标本中均表达编码KGF原型I（即真正的KGF）的mRNA。
