Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, China.
Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu, China; Jiangsu Key Laboratory of Drug Design and Optimization, China Pharmaceutical University, Nanjing, Jiangsu, China.
Biochem Pharmacol. 2015 Jan 15;93(2):210-20. doi: 10.1016/j.bcp.2014.11.010. Epub 2014 Dec 3.
Monolayer cells are largely different from tumor masses, and might misguide drug screenings. 3D in vitro cell culture models simulate the characteristics of tumor masses in vivo and have recently been used in many studies of anti-cancer drugs. Among various 3D cell culture models, multi-cellular layer (MCL) models allow for the direct quantitative assessment of the penetration of chemotherapeutic agents through solid tissue environments without requiring the use of fluorescently labeled drugs or imaging molecules. Therefore, in our present study, a 3D-no base and embedded MCF-7 MCL model was successfully developed for a 14-day culture. Over time, its thickness and cell layers increased and exhibited highly proliferative properties and drug resistance to adriamycin (ADR) with markedly elevated IC50 values. Meanwhile, G2/M stage cycle arrest was also observed, which likely up-regulated P-gp expression through the Chk2/p53/NF-κB pathway. The elevated P-gp expression altered the ADR penetration kinetics in MCF-7 MCLs in vitro by accelerating the apparent penetration of ADR through the intercellular spaces of the MCLs. Additionally, a decreased ADR retention within tumor cells was observed, but could be significantly reversed by a P-gp inhibitor. The attenuated ADR retention in the deeper cells of tumor masses was confirmed in xenografted mice in vivo. This phenomenon could be elucidated by the mathematical modeling of penetration kinetics parameters. Our study provided a new model that evaluated and improved the quantification of the drug penetration kinetics, revealed the relationship between P-gp and drug penetration through tumor masses, and suggested the potential molecular mechanisms.
单层细胞与肿瘤组织有很大的不同，可能会误导药物筛选。3D 体外细胞培养模型模拟了体内肿瘤组织的特征，最近已被广泛应用于许多抗癌药物的研究中。在各种 3D 细胞培养模型中，多细胞层（MCL）模型允许直接定量评估化疗药物通过实体组织环境的渗透，而无需使用荧光标记药物或成像分子。因此，在本研究中，成功建立了一种用于 14 天培养的无基底和嵌入式 MCF-7 MCL 模型。随着时间的推移，其厚度和细胞层增加，并表现出高度增殖特性和对阿霉素（ADR）的耐药性，IC50 值明显升高。同时，也观察到 G2/M 期细胞周期阻滞，这可能通过 Chk2/p53/NF-κB 通路上调 P-糖蛋白（P-gp）的表达。升高的 P-gp 表达改变了 MCF-7 MCL 中 ADR 的渗透动力学，通过加速 ADR 通过 MCL 细胞间空间的表观渗透来实现。此外，还观察到肿瘤细胞内 ADR 的保留减少，但通过 P-gp 抑制剂可显著逆转。在体内异种移植小鼠中证实了肿瘤块深层细胞中 ADR 保留的减少。这种现象可以通过渗透动力学参数的数学建模来解释。本研究提供了一种新的模型，用于评估和改进药物渗透动力学的定量分析，揭示了 P-gp 与药物通过肿瘤组织渗透之间的关系，并提出了潜在的分子机制。
