Irinotecan Synergistically Enhances the Antiproliferative and Proapoptotic Effects of Axitinib In Vitro and Improves Its Anticancer Activity In Vivo
Irinotecan Synergistically Enhances the Antiproliferative and Proapoptotic Effects of Axitinib In Vitro and Improves Its Anticancer Activity In Vivo
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DOI：
10.1593/neo.101334
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发表时间：
2011-03-01
期刊：
影响因子：
4.8
通讯作者：
Bocci, Guido
中科院分区：
文献类型：
--
作者：
Canu, Bastianina;Fioravanti, Anna;Bocci, Guido
目标：证明伊立替康和阿西替尼在体外的协同抗增殖和促凋亡活性，以及对血管生成和胰腺癌的体内作用的改善。方法：对暴露于SN-38、伊立替康活性代谢产物、阿西替尼或其同时组合72小时的人真皮微血管内皮细胞和胰腺癌（MIAPaCa-2、Capan-1）细胞系进行增殖和凋亡试验。采用ELISA法检测ERK 1/2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ATP 7A和ABCG 2基因表达，采用高效液相色谱法测定SN-38细胞内浓度。用伊立替康和阿西替尼单独或同时组合处理裸鼠中的Capan-1异种移植物。研究结果：发现阿西替尼/SN-38组合对内皮细胞和癌细胞的抗增殖和促凋亡活性具有强烈的协同作用。ERK 1/2和Akt磷酸化在所有细胞系中被较低浓度的组合药物显著抑制。阿昔替尼和SN-38联合治疗极大地抑制了内皮细胞和癌细胞中ATP 7A和ABCG 2基因的表达，增加了SN-38的细胞内浓度。此外，TSP-1分泌增加，而VEGFR-2水平显着下降，在细胞与两种药物治疗。体内同时给药确定了肿瘤和肿瘤新血管形成几乎完全消退。结论：体外结果显示，伊立替康和阿昔替尼同时联合对内皮和胰腺癌细胞具有高度协同作用，表明该方案可能转化为临床。
AIMS: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.