Nishimura Masuhiro, Suzuki Satoshi, Satoh Tetsuo, Naito Shinsaku
Research and Development Center, Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan.
Drug Metab Pharmacokinet. 2009;24(1):91-9. doi: 10.2133/dmpk.24.91.
Pairs of forward and reverse primers and TaqMan probes specific to each of 23 human solute carrier 35 (SLC35) transporters were prepared. The mRNA expression level of each target transporter was analyzed in total RNA from single and pooled specimens of adult human tissues (adipose tissue, adrenal gland, bladder, bone marrow, brain, cerebellum, colon, heart, kidney, liver, lung, mammary gland, ovary, pancreas, peripheral leukocytes, placenta, prostate, retina, salivary gland, skeletal muscle, small intestine, smooth muscle, spinal cord, spleen, stomach, testis, thymus, thyroid gland, tonsil, trachea, and uterus), from pooled specimens of fetal human tissues (brain, heart, kidney, liver, spleen, and thymus), and from three human cell lines (HeLa cell line ATCC#: CCL-2, human cell line Hep G2, and human breast carcinoma cell line MDA-435) by real-time reverse transcription PCR using an Applied Biosystems 7500 Fast Real-Time PCR System. The mRNA expression of SLC35As, SLC35Bs, SLC35Cs, SLC35D1, SLC35D2, SLC35Es, and SLC35F5 was found to be ubiquitous in both adult and fetal tissues. SLC35D3 mRNA was expressed at the highest levels in the adult retina. SLC35F1 mRNA was expressed at high levels in the adult and fetal brain. SLC35F2 mRNA was expressed at the highest levels in the adult salivary gland. Both SLC35F3 and SLC35F4 mRNAs were expressed at the highest levels in the adult cerebellum. Further, individual differences in the mRNA expression levels of human SLC35 transporters in the liver were also evaluated. Our newly determined expression profiles were used to study the gene expression in 31 adult human tissues, 6 fetal human tissues, and 3 cell lines, and tissues with high transcriptional activity for human SLC35 transporters were identified. These results are expected to be valuable for research concerning the clinical diagnosis of disease.
制备了针对23种人类溶质载体35(SLC35)转运蛋白的正向和反向引物对以及TaqMan探针。使用Applied Biosystems 7500快速实时PCR系统,通过实时逆转录PCR分析了来自成人人类组织(脂肪组织、肾上腺、膀胱、骨髓、脑、小脑、结肠、心脏、肾脏、肝脏、肺、乳腺、卵巢、胰腺、外周白细胞、胎盘、前列腺、视网膜、唾液腺、骨骼肌、小肠、平滑肌、脊髓、脾脏、胃、睾丸、胸腺、甲状腺、扁桃体、气管和子宫)的单个和混合标本、胎儿人类组织(脑、心脏、肾脏、肝脏、脾脏和胸腺)的混合标本以及三种人类细胞系(HeLa细胞系ATCC#:CCL - 2、人类细胞系Hep G2和人类乳腺癌细胞系MDA - 435)的总RNA中各目标转运蛋白的mRNA表达水平。发现SLC35A、SLC35B、SLC35C、SLC35D1、SLC35D2、SLC35E和SLC35F5的mRNA表达在成人和胎儿组织中均普遍存在。SLC35D3 mRNA在成人视网膜中表达水平最高。SLC35F1 mRNA在成人和胎儿脑中表达水平较高。SLC35F2 mRNA在成人唾液腺中表达水平最高。SLC35F3和SLC35F4 mRNA在成人小脑中表达水平均最高。此外,还评估了人类肝脏中SLC35转运蛋白mRNA表达水平的个体差异。我们新确定的表达谱用于研究31种成人人类组织、6种胎儿人类组织和3种细胞系中的基因表达,并确定了人类SLC35转运蛋白具有高转录活性的组织。这些结果有望对疾病临床诊断的研究具有重要价值。