College of Pharmaceutical Science, Zhejiang University of Technology, No. 18 Chaowang Road, Hangzhou, 310014, China.
Collaborative Innovation Center of Yangtze River Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou, China.
Cancer Chemother Pharmacol. 2019 Sep;84(3):527-537. doi: 10.1007/s00280-019-03846-x. Epub 2019 Apr 27.
Recent researches are attempting to verify that the 3D cell models can provide a gap bridge between in vitro and in vivo for anticancer drug evaluations. The aim of this study was to continue the development of novel in vitro 3D cell models and the investigation of the cellular uptake mechanism of camptothecin (CPT) and its derivatives [FL118 (10,11-methylenedioxy-20(S)-camptothecin), 9-Q20 (9-p-trifluoromethylphenyl-10,11-methylenedioxy-20(S)-camptothecin)] in 2D and 3D cell models.
The 3D cell models were established using ultralow attachment 96-well plates. The cytotoxicity of CPT, FL118, and 9-Q20 was evaluated by MTT method. The effects of compound concentration, incubation time, temperature, and transporter inhibitors on the cellular uptake of CPT, FL118, and 9-Q20 were examined in 2D and 3D cell models.
The cytotoxicity of CPT, FL118, and 9-Q20 was lower in 3D cell models than 2D cell models. In 2D Caco-2 cell model, the uptake rate of CPT, FL118, and 9-Q20 was faster during the early time of incubation. In 3D Caco-2 cell model, the uptake capacity of CPT, FL118, and 9-Q20 was significantly improved over time. In 3D Caco-2 cell model, Verapamil (P-gp inhibitor) and Gefitinib (BCRP inhibitor) more significantly increased the uptake of CPT and 9-Q20. In contrast, P-gp and BCRP did not affect the accumulation of FL118 in 2D and 3D Caco-2 cell models. The accumulation of CPT, FL118, and 9-Q20 was greater in HepG2 cells than HCT116 cells.
The 3D cell models provided more potency information on the process of cellular uptake of CPT, FL118, and 9-Q20, which more objectively reflected the drug sensitivity and drug resistance in vivo compared with the 2D cell models.
最近的研究试图验证 3D 细胞模型可以为抗癌药物评估提供体外和体内之间的间隙桥。本研究的目的是继续开发新型的体外 3D 细胞模型，并研究喜树碱（CPT）及其衍生物[FL118（10,11-亚甲二氧基-20(S)-喜树碱），9-Q20（9-p-三氟甲基苯基-10,11-亚甲二氧基-20(S)-喜树碱）]在 2D 和 3D 细胞模型中的细胞摄取机制。
使用超低附着 96 孔板建立 3D 细胞模型。通过 MTT 法评估 CPT、FL118 和 9-Q20 的细胞毒性。在 2D 和 3D 细胞模型中，研究了化合物浓度、孵育时间、温度和转运体抑制剂对 CPT、FL118 和 9-Q20 细胞摄取的影响。
CPT、FL118 和 9-Q20 在 3D 细胞模型中的细胞毒性低于 2D 细胞模型。在 2D Caco-2 细胞模型中，CPT、FL118 和 9-Q20 的摄取速率在孵育早期较快。在 3D Caco-2 细胞模型中，CPT、FL118 和 9-Q20 的摄取能力随时间显著提高。在 3D Caco-2 细胞模型中，维拉帕米（P-糖蛋白抑制剂）和吉非替尼（BCRP 抑制剂）更显著地增加了 CPT 和 9-Q20 的摄取。相比之下，P-糖蛋白和 BCRP 对 2D 和 3D Caco-2 细胞模型中 FL118 的积累没有影响。CPT、FL118 和 9-Q20 在 HepG2 细胞中的积累量高于 HCT116 细胞。
3D 细胞模型为 CPT、FL118 和 9-Q20 的细胞摄取过程提供了更多的效力信息，与 2D 细胞模型相比，更客观地反映了体内的药物敏感性和耐药性。
