Microarray-based Detection of Multidrug Resistance in Human Tumor Cells by Expression Profiling of ATP-binding Cassette Transporter Genes

Abstract

Different mechanisms of drug resistance, including ATP-binding cassette (ABC) transporters, are responsible for treatment failure of tumors. We developed a low-density DNA microarray which contains 38 genes of the ABC transporter gene family. This tool has been validated with three different multidrug-resistant sublines (CEM/ADR5000, HL60/AR, and MCF7/CH1000) known to overexpress either the ABCB1 (MDR1), ABCC1 (MRP1), or ABCG2 (MXR and BCRP) genes. When compared with their drug-sensitive parental lines, we observed not only the overexpression of these genes in the multidrug-resistant cell lines but also of other ABC transporter genes pointing to their possible role in multidrug resistance. These results were corroborated by quantitative real-time reverse transcription-PCR. As the microarray allows the determination of the expression profile of many ABC transporters in a single hybridization experiment, it may be useful as a diagnostic tool to detect drug resistance in clinical samples.

Use of arrays to investigate the contribution of ATP-binding cassette transporters to drug resistance in cancer chemotherapy and prediction of chemosensitivity

“…Although the most frequently selected ABC transporters in various drug resistant cell lines appeared to be ABCB1 and ABCC1, consistent with their known roles in MDR, new roles in drug resistance previously unknown for some ABC transporters were also found using these profiling analyses. For example, ABCC3 and ABCC5 were both found upregulated in the Adriamycin-selected MCF7/AdVp1000 [24] and MCF7/AdVp3000 [40] cell lines using two different arrays, strongly suggesting that these transporters may contribute to Adriamycin resistance. Although these transporters may be selected by verapamil that was present www.cell-research.com | Cell Research Jian-Ting Zhang 321 npg during the selection of these cell lines, the role of ABCC3 in Adriamycin resistance has been validated functionally by the siRNA knock down strategy [40].…”

“…Eighty-eight such classifiers performed accurately to predict drug sensitivity. Unfortunately, no Table 1 Human ABC Transporters Subfamily Members ABCA ABCA1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A12, A13 ABCB ABCB1, B2, B3, B4, B5, B6, B7, B8, B9, B10, B11 ABCC ABCC1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13 ABCD ABCD1, D2, D3, D4 ABCE ABCE1 ABCF ABCF1, F2, F3 ABCG ABCG1, G2, G4, G5, G8 MCF7/AdVp1000 Adriamycin [24] MCF7/AdVp3000 Adriamycin [40] a The functional validation was conducted following identification by array analyses to verify the functional role of the ABC transporter in resistance to corresponding drugs tested. Some ABC transporters have known roles in resistance to the corresponding drugs from prior studies, which are not counted as functional validation in this table.…”

“…Neither of these two genes has been shown previously to cause drug resistance. However, in the profiling study by Gillet et al using their low density DualChip human ABC microarray (see below), ABCB2 and ABCB9 were also found to be over-expressed in Adriamycin-resistant HL-60 and CCRF-CEM cell lines, respectively [24] (see below). It is currently unknown if the over-expression of ABCB2 and ABCB9 causes Adriamycin resistance.…”

“…Recently, low density arrays containing only ABC transporter genes have been developed which can be used more cost-effectively for focused studies of ABC transporters in drug resistant versus sensitive cells ( Table 2). The first of such arrays contains 38 human ABC transporter genes (DualChip human ABC low-density microarray) and it has been validated using three MDR cell lines, CEM/ADR5000, HL60/AR, and MCF7/AdVp1000 in comparison with their respective parental CEM, HL60, and MCF7 cells [24]. Using this array it was found that CEM/ADR5000, HL60/AR, and MCF7/AdVp1000 over-expressed ABCB1, ABCC1, and ABCG2, respectively, as previously demonstrated [5,31,32], thus validating the usefulness of this low density array.…”

ATP-binding-cassette transporters in hematopoietic stem cells and their utility as therapeutical targets in acute and chronic myeloid leukemia

“…25 Additionally, recent studies examining ABC transporter gene expression in drug-selected cancer cell lines demonstrated overexpression of a number of transporters not previously recognized as associated with drug resistance (ABCA4, ABCA7, ABCB2, ABCB6, ABCB8, ABCB9 and ABCG1). 26 With the exception of ABCC4, all these transporters are expressed in human CD34 þ 38À hematopoietic cells, again many differentially in comparison to committed progenitors. Together, these data outline the emerging picture that ABC-transporter-mediated protection against xenobiotics and other toxic substrates may be conferred by simultaneous activity of many redundant family members in addition to ABCB1, ABCC1 and ABCG2.…”