Studies Of Antiangiogenic Effects And Mechanisms Of Novel Recombinant Human Endostatin Combined With Cisplatin

Objective: To reveal the synergistic antiangiogenic effects and mechanisms of novel recombinant human endostatin (Endostar) combined with cytotoxic drug cisplatin. Methods: On the cultivated system of human umbilical vein endothelial cells (HUVECs), we systematically investigated the antiangiogenic effects of Endostar combined with cisplatin, using the methods including proliferation inhibition assay, clonogenic assay , induction apoptosis, migration/invasion assay, and tube formation study, and in the chicken chorioallantoic membrane (CAM) assay, we observed and evaluated the antiangiogenic properties of combined modality. Meanwhile, we displayed the early gene expression profiles of different therapeutic modalities on HUVECs using DNA microarray technology, and identified some important genes involved in angiogenic modulation. We discussed the roles of these genes contributed to enhanced antiangiogenic effects, and confirmed the gene-chip data using real-time quantitative RT-PCR analysis. Results: There was a synergistic antiangiogenic effect in vitro when Endostar was used with cisplatin, and this synergistic effect was specific on endothelial cells. Endostar had notable antiangiogenic activity, with a nonliner dose-effect relationship, and combined Endostar with cisplatin exhibited a synergistic antiangiogenic effect in vivo. Some important genes associated with enhanced antiangiogenic action, such as JUNB, GATA2, INSR, F7, and WNT1, etc, were identified. The chip data of the target genes were mainly in accordance with the results of real-time quantitative RT-PCR. Conclusions: Combined Endostar with cisplatin resulted in a synergistic antiangiogenic effect, and the molecular mechanism was involved in the significant regulation of some genetic expression. These effectors could enhance the antiangiogenic effect through VEGF/VEGFR, coagulation cascade, and WNT signaling pathways.