P-glycoprotein (P-gp) is an efflux transporter protecting cells from various chemical threats. Upregulation of P-gp expression, mediated by reactive oxygen species (ROS), has been reported in doxorubicin-treated cells. However, during cell differentiation cellular redox status may vary. This study investigated the influence of cell differentiation on doxorubicin-mediated P-gp expression in Caco-2 cells, and its relation to cellular redox status. During 21-day culture, Caco-2 cells were divided into pre-, during-, and post-differentiation, based on total cell numbers, percentage of G0/G1 cells, and alkaline phosphatase activity. Our study demonstrated that Caco-2 cells were in a more reduced state on differentiation. At low concentrations (<5–10 μM), doxorubicin increased ABCB1 mRNA levels in all differentiation states. The inductive effect vanished on increasing doxorubicin concentrations to 50–100 μM. Moreover, N-acetylcysteine inhibited doxorubicin-mediated up-regulation of ABCB1 mRNA in the pre-differentiated and differentiating cells, but not in the post-differentiated cells. Our findings suggested that Caco-2 differentiation states affect doxorubicin-mediated ABCB1 transcription and intracellular ROS level. Cellular redox status may contribute to the ABCB1 transcription through ROS-dependent mechanisms in the pre- and during-differentiation phases, but not in the post-differentiation phase. Furthermore, the differentiation status should be concern for P-gp induction study in Caco-2 cell model.
Faculty of Pharmaceutical Sciences, Chulalongkorn University
Udomnilobol, Udomsak and Jianmongko, Suree
"Doxorubicin modulates P-glycoprotein transcription differently during Caco-2 differentiation,"
The Thai Journal of Pharmaceutical Sciences: Vol. 45:
2, Article 1.
Available at: https://digital.car.chula.ac.th/tjps/vol45/iss2/1