| RRC ID |
6192
|
| 著者 |
Kugawa F, Suzuki T, Miyata M, Tomono K, Tamanoi F.
|
| タイトル |
Construction of a model cell line for the assay of MDR1 (multi drug resistance gene-1) substrates/inhibitors using HeLa cells.
|
| ジャーナル |
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.
|
| 巻・号 |
64(5)
|
| ページ |
296-300
|
| 公開日 |
2009-5-1
|
| PMID |
19530439
|
| PMC |
PMC4370427
|
| MeSH |
ATP Binding Cassette Transporter, Subfamily B
ATP Binding Cassette Transporter, Subfamily B, Member 1 / biosynthesis
ATP Binding Cassette Transporter, Subfamily B, Member 1 / drug effects
ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics*
Cell Line
Fluorescent Dyes
Genetic Vectors
HeLa Cells
Humans
Models, Genetic
Rhodamine 123
Transfection
|
| IF |
1.198
|
| 引用数 |
4
|
|
WOS 分野
|
CHEMISTRY, MULTIDISCIPLINARY
PHARMACOLOGY & PHARMACY
CHEMISTRY, MEDICINAL
|
| リソース情報 |
| 遺伝子材料 |
Human MDR1 cDNA (RDB01372) |
