Topotecan is a substrate for multidrug resistance associated protein 4

Tian, Quan, Zhang, Jing, Chan, Sui Y., Chin Tan, Theresa M., Duan, Wei, Huang, Min, Zhu, Yi Z., Chan, Eli, Yu, Qiang, Nie, Yu Q.i.a.n.g, Chi-Liu Ho, Paul, Li, Qi, Ng, Ka-Yun, Yang, Hong-Yuan, Wei, Hong, Bian, Jin-Song and Zhou, Shu-Feng 2006, Topotecan is a substrate for multidrug resistance associated protein 4, Current drug metabolism, vol. 7, no. 1, pp. 105-118.

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Title Topotecan is a substrate for multidrug resistance associated protein 4
Author(s) Tian, Quan
Zhang, Jing
Chan, Sui Y.
Chin Tan, Theresa M.
Duan, WeiORCID iD for Duan, Wei
Huang, Min
Zhu, Yi Z.
Chan, Eli
Yu, Qiang
Nie, Yu Q.i.a.n.g
Chi-Liu Ho, Paul
Li, Qi
Ng, Ka-Yun
Yang, Hong-Yuan
Wei, Hong
Bian, Jin-Song
Zhou, Shu-Feng
Journal name Current drug metabolism
Volume number 7
Issue number 1
Start page 105
End page 118
Publisher Bentham Science Publishers Ltd
Place of publication Hilversum, Netherlands
Publication date 2006-01
ISSN 1389-2002
Keyword(s) Camptothecin
multidrug resistance associated protein 4
HepG2 cells
Summary Topotecan (TPT) is a semisynthetic water-soluble derivative of camptothecin (CPT) used as second-line therapy in patients with metastatic ovarian carcinoma, small cell lung cancer, and other malignancies. However, both doselimiting toxicity and tumor resistance hinder the clinical use of TPT. The mechanisms for resistance to TPT are not fully defined, but increased efflux of the drug by multiple drug transporters including P-glycoprotein (PgP), multidrug resistance associated protein 1 (MRP1) and breast cancer resistance protein (BCRP) from tumor cells has been highly implicated. This study aimed to investigate whether overexpression of human MRP4 rendered resistance to TPT by examining the cytotoxicity profiles using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazonium bromide (MTT) assay and cellular accumulation of TPT in HepG2 cells stably overexpressing MRP4. Two kinds of cell lines, HepG2 with insertion of an empty vector plasmid (V/HepG2), HepG2 cells stably expressing MRP4 (MRP4/HepG2), were exposed to TPT for 4 or 48 hr in the absence or presence of various MRP4 inhibitors including DL-buthionine-(S,R)-sulphoximine (BSO), diclofenac, celecoxib, or MK-571. The intracellular accumulation of TPT and paclitaxel (a PgP substrate) by V/HepG2 and MRP4/HepG2 cells was determined by incubation of TPT with the cells and the amounts of the drug in cells were determined by validated HPLC methods. The study demonstrated that MRP4 conferred a 12.03- and 6.86-fold resistance to TPT in the 4- and 48-hr drug-exposure MTT assay, respectively. BSO, MK-571, celecoxib, or diclofenac sensitised MRP4/HepG2 cells to TPT cytotoxicity and partially reversed MRP4-mediated resistance to TPT. In addition, the accumulation of TPT was significantly reduced in MRP4/HepG2 cells compared to V/HepG2 cells, and one-binding site model was found the best fit for the MRP4-mediated efflux of TPT, with an estimated Km of 1.66 mM and Vmax of 0.341 ng/min/106 cells. Preincubation of MRP4/HepG2 cells with BSO (200 μM) for 24 hr, celecoxib (50 mM), or MK-571 (100 mM) for 2 hr significantly increased the accumulation of TPT over 10 min in MRP4/HepG2 cells by 28.0%, 37.3% and 32.5% (P < 0.05), respectively. By contrast, there was no significant difference in intracellular accumulation of paclitaxel in V/HepG2 and MRP4/HepG2 cells over 120 min. MRP4 also rendered resistance to adefovir dipivoxil (bis-POMPMEA) and methotrexate, two reported MRP4 substrates. MRP4 did not exhibit any significant resistance to other model drugs including vinblastine, vincristine, etoposide, carboplatin, cyclosporine and paclitaxel in both long (48 hr) and short (4 hr) drug-exposure MTT assays. These findings indicate that MRP4 confers resistance to TPT and TPT is the substrate for MRP4. Further studies are needed to explore the role of MRP4 in resistance to, toxicity and pharmacokinetics of TPT in cancer patients.
Language eng
Field of Research 110106 Medical Biochemistry: Proteins and Peptides (incl Medical Proteomics)
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2006, Bentham Science Publishers Ltd.
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