Passive permeability assay of doxorubicin through model cell membranes under cancerous and normal membrane potential conditions

摘要

Doxorubicin is an anti-cancer drug that is important for breast cancer therapy. In this study, the effects o f the membrane potential of breast cancer cells (- 30 mV) and normal breast epithelial cells (- 60 mV) on doxorubicin (DOX) permeability was studied. To achieve this goal, black lipid membranes (BLMs) as a model cell membrane were formed with DPhPC phospholipids in a single aperture of a Teflon sheet by the Montal and Mueller method. The presence o f the BLM was characterized by capacitive measurements. The measured specific capacitance of 0.6 mu F/cm(2) after applying the Montal and Mueller method, confirming the presence of a BLM in the aperture. In addition, the very low current leakage o f the BLM (9-24 pA) and ClyA-protein channel insertion in the BLM indicate the compactness, high quality, and thickness of 3-5 nm of the BLM. Afterwards, the permeability of doxorubicin through the BLM was studied at defined cell conditions (37 degrees C and pH 7.4), as well as cancerous and healthy epithelial-cell membrane potentials (- 3 0 mV and - 60 mV, respectively). The results show a slow DOX penetration within the first few hours, which increases rapidly with time. The initial slow penetration can be attributed to an electrostatic interaction between doxorubicin and DPhPC molecules in the model cell membrane. Furthermore, a MTT assay on MCF-10A and MCF-7 under different concentrations of doxorubicin confirmed that the cancerous MCF-7 cell line is more resistant to doxorubicin in comparison with the non-cancerous MCF-10A. Such studies highlight important strategies for designing and tuning the interaction efficacy of novel pharmaceuticals at molecular level.