RRC ID 6192
Author Kugawa F, Suzuki T, Miyata M, Tomono K, Tamanoi F.
Title Construction of a model cell line for the assay of MDR1 (multi drug resistance gene-1) substrates/inhibitors using HeLa cells.
Journal Pharmazie
Abstract Cancer cells often become resistant to chemotherapy, and induction of the ABC transporter Multi-drug Resistance gene-1 (MDR1) is a major cause. We established a tool for high-throughput screening of substrates and inhibitors of MDR1, using transformed HeLa cells that over-express MDR1. The cDNA for human MDR1 was subcloned into the eukaryotic expression vector pBK-CMV to produce an MDR1 expression vector, pBK-CMV/MDR1. HeLa cells were transfected with pBK-CMV/MDR1 or the empty vector pBK-CMV. Transfection of the vector sequence for MDR1 and its expression were evaluated by genomic PCR and western blotting, respectively. The efficiency of the MDR1 transporter for pumping a substrate out of the transformed cells was evaluated using rhodamine123 (R-123), a mitochondrial dye that is also an MDR1 substrate. After treatment of the MDR1-expressing HeLa cells with MDR1 substrate vinblastin or inhibitors cyclosporin A and verapamil, the amount of R-123 retained in the cells was increased to 2 to 2.3 times the level in untreated MDR1-expressing HeLa cells. The transfection of empty pBK-CMV had no effect on the R-123 retention in HeLa cells, regardless of drug treatment. In conclusion, we have established a model human carcinoma cell line that expresses functional MDR1 and can be used to screen for substrates and inhibitors of MDR1.
Volume 64(5)
Pages 296-300
Published 2009-5
PMID 19530439
PMC PMC4370427
MeSH ATP Binding Cassette Transporter, Sub-Family B ATP-Binding Cassette, Sub-Family B, Member 1 / biosynthesis ATP-Binding Cassette, Sub-Family B, Member 1 / drug effects ATP-Binding Cassette, Sub-Family B, Member 1 / genetics* Cell Line Fluorescent Dyes Genetic Vectors HeLa Cells Humans Models, Genetic Rhodamine 123 Transfection
IF 1.126
Times Cited 4
WOS Category CHEMISTRY, MULTIDISCIPLINARY PHARMACOLOGY & PHARMACY CHEMISTRY, MEDICINAL
Resource
DNA material Human MDR1 cDNA (RDB01372)