Trebinska-Stryjewska Alicja, Swiech Olga, Opuchlik Lidia J, Grzybowska Ewa A, Bilewicz Renata
Institute of Optoelectronics, Biomedical Engineering Centre, Military University of Technology, 00-908 Warsaw, Poland.
Department of Molecular and Translational Oncology, Maria Sklodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland.
ACS Omega. 2020 Apr 1;5(14):7979-7986. doi: 10.1021/acsomega.9b04479. eCollection 2020 Apr 14.
The influence of the pH of the multicomponent cell medium on the performance of doxorubicin (DOX), an anticancer drug, was studied on the examples of cervical (HeLa) and kidney (A498) cancer cell lines. The change of pH of the cell medium to more acidic led to a decrease of DOX toxicity on both cell lines due to the change of drug permeability across the cell membrane as a result of drug protonation. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) studies and lactate dehydrogenase (LDH) release tests have shown low toxicity of the drug, especially in the case of A498 cells, which are characterized by an extremely high glycolytic metabolism. The behavior was ascribed primarily to the increased proton concentration in the peripheral blood follicle in the presence of products of the acidic glycolytic metabolism. It is not observed in the measurements performed in commercially available media since they usually have a neutral pH. In earlier reports on kidney cancer, several mechanisms were discussed, including the metabolism of DOX to its less toxic derivative, doxorubicinol, overexpression of ATP binding cassette subfamily B member 1 (ABCB1) transporters, that remove DOX from the inside of cells; however, there was no focus on the simple but very important contribution of drug protonation described in the present study. Drug pH-dependent equilibria in the cell medium should be considered since changes in the drug form may be an additional reason for multidrug resistance.
以宫颈癌细胞系（HeLa）和肾癌细胞系（A498）为例，研究了多组分细胞培养基的pH值对抗癌药物阿霉素（DOX）性能的影响。细胞培养基pH值向更酸性的变化导致两种细胞系上DOX毒性降低，这是由于药物质子化导致跨细胞膜的药物通透性发生变化。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）研究和乳酸脱氢酶（LDH）释放试验表明该药物毒性较低，尤其是在具有极高糖酵解代谢特征的A498细胞中。这种行为主要归因于在酸性糖酵解代谢产物存在的情况下外周血卵泡中质子浓度增加。在市售培养基中进行的测量未观察到这种情况，因为它们通常具有中性pH值。在早期关于肾癌的报道中，讨论了几种机制，包括DOX代谢为毒性较小的衍生物阿霉素醇、ATP结合盒亚家族B成员1（ABCB1）转运蛋白的过表达，该转运蛋白可将DOX从细胞内清除；然而，本研究中描述的药物质子化这一简单但非常重要的作用并未受到关注。应考虑细胞培养基中药物的pH依赖性平衡，因为药物形式的变化可能是多药耐药的另一个原因。
