Real-time reverse transcription-PCR expression profiling of the complete human ATP-binding cassette transporter superfamily in various tissues
Real-time reverse transcription-PCR expression profiling of the complete human ATP-binding cassette transporter superfamily in various tissues
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DOI：
10.1373/49.2.230
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发表时间：
2003-02-01
影响因子：
9.3
通讯作者：
Schmitz, G
中科院分区：
文献类型：
--
作者：
Langmann, T;Mauerer, R;Schmitz, G
背景。atp结合盒（ABC）转运体参与许多生理过程，如脂质转运、固醇稳态、免疫机制和药物转运，并引起各种人类遗传性疾病。因此，分析ABC转运蛋白mRNA的表达谱对基础研究，特别是在脂质代谢、临床诊断和药物作用监测等领域具有重要意义。我们开发了一种快速、准确、高灵敏度的实时逆转录pcr （RT-PCR）方法，用于检测和定量目前已知的ABC转运蛋白超家族的所有47个成员。我们的表达分析是基于使用校准曲线方法的相对定量。通过我们的分析，可以在384孔格式中监测大量RNA样品的表达，总RNA仅为50 ng。结果：与以往单个ABC基因的表达分析相比，我们的方法可以快速完整地分析给定RNA样本中的所有ABC转运蛋白。我们使用新建立的表达面板研究了所有人类ABC转运蛋白在20种不同人体组织中的基因表达。因此，我们确定了ABC转运蛋白具有高转录活性的组织。这些器官主要涉及分泌功能（肾上腺）、代谢功能（肝脏）、屏障功能（肺、气管、小肠）和回归线功能（胎盘、子宫）。结论：我们的RT-PCR方法可以快速、高通量地分析ABC转运蛋白超家族，从而为研究、疾病临床诊断、药物测试和开发提供新的支持。(C) 2003年美国临床化学学会。
Background. ATP-binding cassette (ABC) transporters are involved in many physiologic processes, such as lipid transport, sterol homeostasis, immune mechanisms, and drug transport, and cause various human inherited diseases. Thus, the analysis of ABC transporter mRNA expression profiles for basic research, especially in the field of lipid metabolism, for clinical diagnosis, and for monitoring of drug effects is of great interest.Methods. We have developed a rapid, accurate, and highly sensitive real-time reverse transcription-PCR (RT-PCR) method for detection and quantification of all 47 currently known members of the ABC transporter superfamily. Our expression analysis is based on relative quantification using a calibration curve method. With our assay, expression monitoring of a large number of RNA samples in a 384-well format with only 50 ng of total RNA is possible.Results: In contrast to previous expression analyses of single ABC genes, our method allows the rapid and complete analysis of all ABC transporters in given RNA samples. We used our newly established expression panel to study the gene expression of all human ABC transporters in 20 different human tissues. As a result, we identified tissues with high transcriptional activity for ABC transporters. These organs are mainly involved in secretory function (adrenal gland), metabolic function (liver), barrier function (lung, trachea, small intestine), and tropic function (placenta, uterus).Conclusions: Our RT-PCR assay allows rapid, high-throughput transcriptional profiling of the complete ABC transporter superfamily and thus provides a new enabling toot for research, clinical diagnosis of disease, and drug testing and development. (C) 2003 American Association for Clinical Chemistry.