Individual differences in the response to fentanyl, which may be caused by different concentrations of the drug in the central nervous system, can complicate analgesic treatment. It has been reported that the organic anion transporting polypeptide (OATP) at the blood‑brain barrier (BBB) in Sprague‑Dawley rats may serve an important role in the transport of fentanyl across the BBB. However, whether human OATP can transport fentanyl has thus far not been reported. The present study aimed to establish a 293 cell line stably overexpressing OATP1A2, and to determine whether OATP1A2 is able to transport fentanyl across the plasma membrane. Initially, 293 cells were transfected with an OATP1A2‑expressing plasmid (referred to as 293‑OATP1A2 cells), and single colonies were selected and characterized following geneticin treatment. Subsequently, reverse transcription‑quantitative polymerase chain reaction and western blot analyses were conducted to verify the transfection efficiency. Furthermore, treatment of 293‑OATP1A2 cells with different concentrations of fexofenadine (FEX) and fentanyl was performed to investigate the transport function of OATP1A2 in 293 cells. FEX and fentanyl uptake experiments were also performed with naringenin, an inhibitor of OATP1A2. The results indicated that FEX and fentanyl uptake was significantly increased in 293‑OATP1A2 cells compared with that in the control‑transfected cells. The 293‑OATP1A2‑mediated uptake of FEX at concentration of 100 nM FEX was ~10‑fold higher than that of 293‑VC cells. The 293‑OATP1A2‑mediated uptake of fentanyl (100 nM) was 5.1‑fold higher compared with that in 293‑VC cells. In 293‑OATP1A2 cells, the uptake of FEX without OATP1A2 inhibitor naringenin (100 µg/ml) was 2.8‑fold higher compared with that in the presence of naringenin, and the uptake of fentanyl without naringenin was 7.3‑fold higher compared with that in the presence of naringenin (100 µg/ml). In conclusion, 293 cells that overexpressed OATP1A2 were successfully constructed, and OATP1A2 was revealed to mediate fentanyl uptake in the cultured cells.
Keywords: organic anion transporting polypeptide 1a2; fentanyl; substrate; transport function; overexpression.