Division of Pharmacology, Department of Internal Medicine, University of Pisa, Pisa, Italy.
Neoplasia. 2011 Mar;13(3):217-29. doi: 10.1593/neo.101334.
To demonstrate the synergistic antiproliferative and proapoptotic activity of irinotecan and axitinib in vitro and the improvement of the in vivo effects on angiogenesis and pancreatic cancer.
Proliferation and apoptotic assays were performed on human dermal microvascular endothelial cells and pancreas cancer (MIAPaCa-2, Capan-1) cell lines exposed to SN-38, the active metabolite of irinotecan, axitinib, or their simultaneous combination for 72 hours. ERK1/2 and Akt phosphorylation, the vascular endothelial growth factor (VEGF), VEGF receptor-2, and thrombospondin-1 (TSP-1) concentration were measured by ELISAs. ATP7A and ABCG2 gene expression was performed with real-time polymerase chain reaction and SN-38 intracellular concentrations were measured by high-performance liquid chromatography. Capan-1 xenografts in nude mice were treated with irinotecan and axitinib alone or in simultaneous combination.
A strong synergistic effect on antiproliferative and proapoptotic activity was found with the axitinib/SN-38 combination on endothelial and cancer cells. ERK1/2 and Akt phosphorylation were significantly inhibited by lower concentrations of the combined drugs in all the cell lines. Axitinib and SN-38 combined treatment greatly inhibited the expression of the ATP7A and ABCG2 genes in endothelial and cancer cells, increasing the SN-38 intracellular concentration. Moreover, TSP-1 secretion was increased in cells treated with both drugs, whereas VEGFR-2 levels significantly decreased. In vivo administration of the simultaneous combination determined an almost complete regression of tumors and tumor neovascularization.
In vitro results show the highly synergistic properties of simultaneous combination of irinotecan and axitinib on endothelial and pancreas cancer cells, suggesting a possible translation of this schedule into the clinics.
展示伊立替康活性代谢物 SN-38 与阿昔替尼联合应用在体外的协同抗增殖和促凋亡活性，以及改善对血管生成和胰腺癌的体内作用。
对人真皮微血管内皮细胞和胰腺癌细胞（MIAPaCa-2、Capan-1）系进行增殖和凋亡检测，使其暴露于 SN-38（伊立替康的活性代谢物）、阿昔替尼或二者同时联合应用 72 小时。采用 ELISA 法检测 ERK1/2 和 Akt 磷酸化、血管内皮生长因子（VEGF）、VEGF 受体-2 和血小板反应蛋白-1（TSP-1）浓度。采用实时聚合酶链反应检测 ATP7A 和 ABCG2 基因表达，采用高效液相色谱法测量 SN-38 细胞内浓度。将 Capan-1 异种移植瘤裸鼠用伊立替康和阿昔替尼单独或同时联合治疗。
在血管内皮细胞和癌细胞中，阿昔替尼/SN-38 联合应用具有很强的协同抗增殖和促凋亡作用。在所有细胞系中，较低浓度的联合药物显著抑制 ERK1/2 和 Akt 磷酸化。阿昔替尼和 SN-38 联合治疗可显著抑制内皮细胞和癌细胞中 ATP7A 和 ABCG2 基因的表达，增加 SN-38 细胞内浓度。此外，两种药物处理的细胞中 TSP-1 分泌增加，而 VEGFR-2 水平显著降低。同时联合给药的体内给药导致肿瘤和肿瘤新生血管几乎完全消退。
体外结果表明伊立替康和阿昔替尼同时联合应用对血管内皮细胞和胰腺癌细胞具有高度协同作用，提示该方案可能转化为临床应用。
