Biomedical Engineering Faculty, Amirkabir University of Technology, 424 Hafez Ave, Tehran, Iran; Process Engineering for Sustainable Systems, Department of Chemical Engineering, KU Leuven, Celestijnenlaan 200 F, 3001 Leuven, Belgium; Laboratory for Soft Matter and Biophysics, Department of Physics and Astronomy, KU Leuven, Celestijnenlaan 200 D, 3001 Leuven, Belgium.
Laboratory for Soft Matter and Biophysics, Department of Physics and Astronomy, KU Leuven, Celestijnenlaan 200 D, 3001 Leuven, Belgium.
Eur J Pharm Biopharm. 2020 Jan;146:133-142. doi: 10.1016/j.ejpb.2019.10.011. Epub 2019 Nov 5.
Doxorubicin is an anti-cancer drug that is important for breast cancer therapy. In this study, the effects of 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 of the BLM was characterized by capacitive measurements. The measured specific capacitance of 0.6 µF/cm after applying the Montal and Mueller method, confirming the presence of a BLM in the aperture. In addition, the very low current leakage of 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 °C and pH 7.4), as well as cancerous and healthy epithelial-cell membrane potentials (-30 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.
阿霉素是一种抗癌药物，对乳腺癌治疗很重要。在这项研究中，研究了乳腺癌细胞（-30 mV）和正常乳腺上皮细胞（-60 mV）的膜电位对阿霉素（DOX）通透性的影响。为了实现这一目标，采用 Montal 和 Mueller 方法，用 DPhPC 磷脂在聚四氟乙烯片的单个孔中形成黑脂膜（BLM）作为模型细胞膜。通过电容测量来表征 BLM 的存在。应用 Montal 和 Mueller 方法后，测量到的特定电容为 0.6 µF/cm，证实了孔中存在 BLM。此外，BLM 的极低漏电流（9-24 pA）和 ClyA 蛋白通道在 BLM 中的插入表明 BLM 的紧凑性、高质量和 3-5 nm 的厚度。之后，在定义的细胞条件（37°C 和 pH 7.4）下以及癌细胞和健康上皮细胞膜电位（分别为-30 mV 和-60 mV）下研究了阿霉素通过 BLM 的渗透性。结果表明，DOX 在最初的几个小时内渗透缓慢，随着时间的推移迅速增加。初始缓慢渗透可归因于模型细胞膜中阿霉素和 DPhPC 分子之间的静电相互作用。此外，在不同浓度的阿霉素下对 MCF-10A 和 MCF-7 进行的 MTT 测定证实，与非癌细胞 MCF-10A 相比，癌细胞 MCF-7 系对阿霉素的耐药性更强。这些研究强调了在分子水平上设计和调整新型药物相互作用功效的重要策略。
