Advances in the molecular detection of ABC transporters involved in multidrug resistance in cancer

Abstract

ATP-Binding Cassette (ABC) transporters are important mediators of multidrug resistance (MDR) in patients with cancer. Although their role in MDR has been extensively studied in vitro, their value in predicting response to chemotherapy has yet to be fully determined. Establishing a molecular diagnostic assay dedicated to the quantitation of ABC transporter genes is therefore critical to investigate their involvement in clinical MDR. In this article, we provide an overview of the methodologies that have been applied to analyze the mRNA expression levels of ABC transporters, by describing the technology, its pros and cons, and the experimental protocols that have been followed. We also discuss recent studies performed in our laboratory that assess the ability of the currently available high-throughput gene expression profiling platforms to discriminate between highly homologous genes. This work led to the conclusion that high-throughput TaqMan-based qRT-PCR platforms provide standardized clinical assays for the molecular detection of ABC transporters and other families of highly homologous MDR-linked genes encoding, for example, the uptake transporters (solute carriers-SLCs) and the phase I and II metabolism enzymes.

Figure 1. Correlation of ABCB1 and ABCG2 expression profiles from three distinct platforms

Expression profiles for ABCB1 across all 60 cell lines were compared between: (A) SYBR Green and microarray; (B) TLDA and microarray; (C) TLDA and SYBR Green. Identical comparisons were performed for ABCG2 expression profiles: (D) SYBR Green and microarray; (E) TLDA and microarray; (F) TLDA and SYBR Green. The data show that TLDA provides more sensitivity, yielding a larger dynamic range of measurement. The coefficient of correlation is given for each comparison. Values for both the X and Y axes are normalized intensity data and presented in log base 2. Reprinted from Orina et al. 2009, Mol Cancer Ther 8: 2057–2066.

Figure 2. Correlation of ABCB1, ABCC1 and ABCG2 gene expression profiles obtained from the TLDA and BioMark 48.48 Dynamic Array

Expression profiles for ABCB1 (A), -C1 (B) and -G2 (C) across all NCI-60 cancer cell lines were compared between TLDA and BioMark platforms. Values for both the X and Y axes are normalized intensity data and presented in log base 2. Reprinted from Orina et al. 2009, Mol Cancer Ther 8: 2057–2066.