Effect of hr-IL2 treatment on intestinal P-glycoprotein expression and activity in Caco-2 cells.
Caco-2 cells were used to investigate the effect of human recombinant interleukin-2 (IL2) on intestinal P-glycoprotein (P-gp) transporter activity in-vitro. More specifically the efflux function of P-gp was studied by measuring the transepithelial transport of rhodamine-123, a fluorescent substrate of P-gp. Its transport was completely inhibited by two specific P-gp inhibitors, ciclosporin A and GG918, in our experiments. Conversely, these two specific P-gp inhibitors inhibited only 50% of transepithelial transport when [3H]vincristine was used as substrate. After Caco-2 cells were treated with 100 IU mL-1 (6.1 ng mL-1) IL2 for 24 h, a significant diminution (21%) of P-gp transporter function was observed with rhodamine-123 substrate. This effect was also confirmed after 48 and 72 h of exposure to IL2. However, for higher concentrations of IL2 (1000 and 5000 IU mL-1), diminution of P-gp function only occurred after a longer treatment period (48 h and more). The inhibitory effect of IL2 on P-gp activity was found to be independent of tight junction function as demonstrated by constant transepithelial electrical resistance (TEER) measurements for all experimental conditions encountered in this study (time and concentration of IL2 exposure). Furthermore, the MDR1 mRNA level was found to be strongly repressed in Caco-2 cells exposed with 1000 IU mL-1 IL2 for 72 h while the amount of MRP1 mRNA remained unchanged. In conclusion, acute incubation of Caco-2 cells with IL2 induced a decrease of P-gp transporter expression and activity.
Belliard
Anne-Marie
AM
Laboratoire de Physiologie-Pharmacie Clinique, UPRES 2706, Faculté de Pharmacie, 92296 Châtenay-Malabry, France.
Tardivel
Sylviane
S
Farinotti
Robert
R
Lacour
Bernard
B
Leroy
Christine
C
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis* / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001692:Biological Transport; D018938:Caco-2 Cells; D004068:Digestive System Physiological Phenomena; D004848:Epithelium / Q000502:physiology; D005456:Fluorescent Dyes; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology*; D012313:RNA / Q000032:analysis; D020112:Rhodamine 123
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D005456:Fluorescent Dyes; D007376:Interleukin-2; D020112:Rhodamine 123; D012313:RNA