Nishimura Masuhiro, Naito Shinsaku
Division of Pharmacology, Drug Safety and Metabolism, Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan.
Drug Metab Pharmacokinet. 2005 Dec;20(6):452-77. doi: 10.2133/dmpk.20.452.
Pairs of forward and reverse primers and TaqMan probes specific to each of 46 human ATP-binding cassette (ABC) transporters and 108 human solute carrier (SLC) transporters were prepared. The mRNA expression level of each target transporter was analyzed in total RNA from single and pooled specimens of various human tissues (adrenal gland, bone marrow, brain, colon, heart, kidney, liver, lung, pancreas, peripheral leukocytes, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thymus, thyroid gland, trachea, and uterus) by real-time reverse transcription PCR using an ABI PRISM 7700 sequence detector system. In contrast to previous methods for analyzing the mRNA expression of single ABC and SLC genes such as Northern blotting, our method allowed us to perform sensitive, semiautomatic, rapid, and complete analysis of ABC and SLC transporters in total RNA samples. Our newly determined expression profiles were then used to study the gene expression in 23 different human tissues, and tissues with high transcriptional activity for human ABC and SLC transporters were identified. These results are expected to be valuable for establishing drug transport-mediated screening systems for new chemical entities in new drug development and for research concerning the clinical diagnosis of disease.
制备了针对46种人类ATP结合盒（ABC）转运蛋白和108种人类溶质载体（SLC）转运蛋白的正向和反向引物对以及TaqMan探针。使用ABI PRISM 7700序列检测系统，通过实时逆转录PCR分析来自各种人类组织（肾上腺、骨髓、脑、结肠、心脏、肾脏、肝脏、肺、胰腺、外周血白细胞、胎盘、前列腺、唾液腺、骨骼肌、小肠、脊髓、脾脏、胃、睾丸、胸腺、甲状腺、气管和子宫）的单个和混合样本的总RNA中各目标转运蛋白的mRNA表达水平。与先前分析单个ABC和SLC基因mRNA表达的方法（如Northern印迹法）不同，我们的方法使我们能够对总RNA样本中的ABC和SLC转运蛋白进行灵敏、半自动、快速且完整的分析。然后，我们新确定的表达谱被用于研究23种不同人类组织中的基因表达，并鉴定出对人类ABC和SLC转运蛋白具有高转录活性的组织。这些结果有望对建立新药开发中新型化学实体的药物转运介导筛选系统以及疾病临床诊断研究具有重要价值。
