Nishida K, Sotozono C, Adachi W, Yamamoto S, Yokoi N, Kinoshita S
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Japan.
Curr Eye Res. 1995 Mar;14(3):235-41. doi: 10.3109/02713689509033520.
The expression of TGF-beta 1, TGF-beta 2 and TGF-beta 3 precursors mRNA transcripts in in vivo human corneal cells were studied. Complementary DNA (cDNA) was generated from poly A+RNA extracted from in vivo human corneal epithelial cells, stromal keratocytes and endothelial cells. With the cDNAs as template, polymerase chain reaction (PCR) was carried out using specific primers of TGF-beta 1, TGF-beta 2 and TGF-beta 3 precursors, synthesized by choosing specific nucleotide sequences in the latency-associated peptide region of each precursor. Southern blot analysis of the PCR products was carried out. In corneal epithelial cells, TGF-beta 2 mRNA transcript was strongly expressed; TGF-beta 1 and -beta 3 mRNA transcripts were also expressed; stromal keratocyte samples expressed TGF-beta 1 and -beta 2 mRNA transcripts but not TGF-beta 3 mRNA transcript. Endothelial cells expressed all three transcripts. The present study, together with the authors' previous immunohistochemical study demonstrates that both protein and mRNA of TGF-beta 2-LAP are present in human corneal epithelium, indicating that TGF-beta 2 may play a crucial role in corneal epithelial cell layers.
研究了体内人角膜细胞中转化生长因子β1(TGF-β1)、转化生长因子β2(TGF-β2)和转化生长因子β3(TGF-β3)前体mRNA转录物的表达。从体内人角膜上皮细胞、基质角膜细胞和内皮细胞中提取的聚腺苷酸加尾RNA(poly A+RNA)生成互补DNA(cDNA)。以这些cDNA为模板,使用通过在每种前体的潜伏相关肽区域选择特定核苷酸序列合成的TGF-β1、TGF-β2和TGF-β3前体的特异性引物进行聚合酶链反应(PCR)。对PCR产物进行Southern印迹分析。在角膜上皮细胞中,TGF-β2 mRNA转录物强烈表达;TGF-β1和-β3 mRNA转录物也有表达;基质角膜细胞样本表达TGF-β1和-β2 mRNA转录物,但不表达TGF-β3 mRNA转录物。内皮细胞表达所有三种转录物。本研究与作者先前的免疫组织化学研究一起表明,TGF-β2潜伏相关肽(TGF-β2-LAP)的蛋白质和mRNA都存在于人角膜上皮中,表明TGF-β2可能在角膜上皮细胞层中起关键作用。