Wen Y, Li G W, Bekhor I
Laboratory of Lens Molecular Biology, Doheny Eye Institute and School of Dentistry, University of Southern California, Los Angeles 90033, USA.
Curr Eye Res. 1996 Feb;15(2):125-30. doi: 10.3109/02713689608997404.
A lens epithelial (LE) cell cDNA clone, designated Clone 156, was isolated from a mature rat LE cDNA library by methods of subtractive hybridization. The cDNA sequence of Clone 156 was 521 nucleotides in length, excluding the poly(T)-tail, and it encoded an open reading frame of 115 amino acids. The translated protein shared extensive sequence similarities with macrophage migration inhibitory factor (MIF) from mouse lens and human T-cell lymphocytes. Northern blot hybridization showed that rat lens MIF mRNA is about 500 nucleotides in length and that its expression in mature rat lens is relatively low in comparison with that in other rat tissues. The expression of MIF mRNA in LE of normal rats and of rats treated by feeding a diet of 50% (w/w) galactose was studied by quantitative RT-PCR. The results showed that the expression of MIF mRNA in a 20-day galactosemic rat LE increased twelvefold as compared to that found in control LE. From the results of this study and from what we know about the locale of epithelial cell differentiation in mature rat lenses, it is being proposed that the increase in abundance of MIF mRNA in the cataractous rat lens is correlated with the enhanced proliferation of the undifferentiated epithelial cells.
通过消减杂交方法,从成熟大鼠晶状体上皮(LE)细胞cDNA文库中分离出一个名为克隆156的晶状体上皮细胞cDNA克隆。克隆156的cDNA序列长度为521个核苷酸(不包括聚腺苷酸尾),编码一个115个氨基酸的开放阅读框。翻译后的蛋白质与来自小鼠晶状体和人T细胞淋巴细胞的巨噬细胞迁移抑制因子(MIF)具有广泛的序列相似性。Northern印迹杂交显示,大鼠晶状体MIF mRNA长度约为500个核苷酸,与其他大鼠组织相比,其在成熟大鼠晶状体中的表达相对较低。通过定量逆转录聚合酶链反应(RT-PCR)研究了正常大鼠和喂食50%(w/w)半乳糖饮食处理的大鼠晶状体上皮细胞中MIF mRNA的表达。结果显示,与对照晶状体上皮细胞相比,20天半乳糖血症大鼠晶状体上皮细胞中MIF mRNA的表达增加了12倍。根据本研究结果以及我们对成熟大鼠晶状体上皮细胞分化位置的了解,有人提出白内障大鼠晶状体中MIF mRNA丰度的增加与未分化上皮细胞增殖增强有关。