Analysis of Expression of Drug Resistance-Linked ABC Transporters in Cancer Cells by Quantitative RT-PCR
Membrane Transporters in Drug Discovery and Development, 637, Kapitel 6, Humana Press, Totowa, NJ, (2010)DOI: 10.1007/978-1-60761-700-6\_6
Zusammenfassung
Quantitative real-time PCR (qRT-PCR) boasts many advantages over microarrays. For instance, very low amounts of total RNA are required to yield highly accurate and reproducible detection of transcript levels. As a consequence, qRT-PCR has become a popular technique for assessing gene expression levels and is now the gold standard. In this chapter, qRT-PCR using two distinct chemistries, SYBR Green and TaqMan, are described. We compare ABC transporter levels in various drug-resistant cancer cell lines by employing each method. SYBR Green yields reproducible results; nevertheless, TaqMan chemistry is superior to SYBR Green, as it displays higher specificity and sensitivity. Gene expression analysis by qRT-PCR is a powerful technique and shows potential as a diagnostic tool for predicting drug response in cancer patients.