Expression and activity of the efflux transporters ABCB1, ABCC2 and ABCG2 in the human colorectal carcinoma cell line LS513.
The human colorectal carcinoma cell line LS513 exhibits epithelial morphology, adherent properties and can grow subcutaneously to form tumors in nude mice. Thus, it is a potential model for mouse xenograft efficacy studies. The present study characterized the expression and activity of P-gp, BCRP and MRP2 in LS513 cells. We investigated the expression of these ATP-binding cassette transporters by Western blot and their activity was also examined using cell culture inserts, where the LS513 cells were grown to confluence for 9 days. The transport of model substrates of P-gp (amprenavir, ritonavir and topotecan), BCRP (topotecan) and MRP2 (SN-38) was studied in the apical to basolateral (A-B) and basolateral to apical (B-A) directions. P-gp, BCRP and MRP2 could be detected by western blot. The LS513 cells exhibited markedly higher transport in the B-A direction than in the A-B direction for the probe substrates tested, with efflux ratios (ERs; B-A/A-B) of 10, 21, 40 and 50 for amprenavir, ritonavir, topotecan and SN38, respectively. The ER could be significantly reduced with the addition of inhibitors of P-gp (GF120918), BCRP (FTC), and MRP2 (MK571), confirming the activity of these transporters in the LS513 cells.
Salphati
Laurent
L
Drug Metabolism and Pharmacokinetics Department, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, United States. salphati.laurent@gene.com
Plise
Emile G
EG
Li
Guangmin
G
D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics*; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics*; D000199:Actins / Q000096:biosynthesis; D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents; D051379:Mice; D018613:Microscopy, Confocal; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics*; D013050:Spectrometry, Fluorescence
D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000199:Actins; D000970:Antineoplastic Agents; D007202:Indicators and Reagents; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009363:Neoplasm Proteins; C513055:ABCB1 protein, human; C000715087:ABCC2 protein, human; C116614:ABCG2 protein, human