Li J, Ding S F, Habib N A, Fermor B F, Wood C B, Gilmour R S
Department of Molecular and Cellular Physiology, Institute of Animal Physiology, Babraham, Cambridge, UK.
Int J Cancer. 1994 May 1;57(3):348-52. doi: 10.1002/ijc.2910570310.
A segment of 712 bases coding for part of the human stearoyl-CoA desaturase gene was made by polymerase chain reaction (PCR) using primers based on published rat cDNA sequences. The human PCR product was confirmed by DNA sequencing. It was next cloned into a vector from which anti-sense, highly radioactive RNA transcripts were made in vitro using T7 polymerase. The transcripts were used to probe desaturase mRNA in a number of human tumour and control tissues, using a very sensitive solution hybridization/RNase protection assay. Increased desaturase mRNA levels were found in colonic and oesophageal carcinomas and in hepatocellular adenoma; however, no consistent trend was seen in hepatocellular carcinoma. It is suggested that certain classes of tumour may exhibit increased levels of desaturase mRNA.
利用基于已发表的大鼠cDNA序列设计的引物,通过聚合酶链反应(PCR)制备了一段编码人硬脂酰辅酶A去饱和酶基因部分序列的712个碱基片段。通过DNA测序确认了该人源PCR产物。接下来将其克隆到一个载体中,利用T7聚合酶在体外制备反义、高放射性RNA转录本。使用非常灵敏的溶液杂交/核糖核酸酶保护试验,这些转录本被用于探测多种人类肿瘤组织和对照组织中的去饱和酶mRNA。在结肠癌、食管癌和肝细胞腺瘤中发现去饱和酶mRNA水平升高;然而,在肝细胞癌中未观察到一致的趋势。提示某些类型的肿瘤可能表现出去饱和酶mRNA水平升高。