RT Journal Article
SR Electronic
T1 Demonstration of the Activity of P-glycoprotein by a Semi-automated Fluorometric Method
JF Anticancer Research
JO Anticancer Res
FD International Institute of Anticancer Research
SP 2173
OP 2177
VO 29
IS 6
A1 GABRIELLA SPENGLER
A1 MIGUEL VIVEIROS
A1 MARTA MARTINS
A1 LILIANA RODRIGUES
A1 ANA MARTINS
A1 JOZSEF MOLNAR
A1 ISABEL COUTO
A1 LEONARD AMARAL
YR 2009
UL http://ar.iiarjournals.org/content/29/6/2173.abstract
AB Background: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene. Materials and Methods: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene™ 3000 (Corbett Research) under different conditions. Results: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB. Conclusion: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.