The Role Of ET-1in CD133+Ovarian Cancer Stem Cells Associated Neovascularization

Objective: The purpose of the research was to isolate and identifycancer stem cells-like cells(CSC-LCS) and define their biologicalcharacteristics used the human epithelial ovarian carcinoma cell line3AOas model system. Then, we investigated the mechanism that ET-1contributed to CSC-associated angiogenesis.Methods: CD133+cells were isolated from3AO by immunomagneticbeads (miniMACS). Cell counting was taken to reflect the proliferationability. The self-renew capacity of CD133+cells was detected by flowcytometry (FCM). No more than100CDD133+cells were inoculatedsubcutaneously in nude mice, formation of xenografts demonstrated thisgroup of cells were able to initiate and maintain the heterogeneous tumor.Besides, we also characterized the sensitivity of CDD133+cells againstchemotherapeutic drug. Secondly, after treatment with ET-1, the expressionof vascular endothelial growth factor (VEGF) was evaluated by RT-PCRand western blot to explore whether CD133+cells would make response forangiogenesis. Results:Here, we have obtained CDD133+cancer stem cells from thehuman epithelial ovarian carcinoma cell line3AO, which are capable ofself-renew, proliferation and differentiated to a number of CDD133-cells.In addition, nude mice xenograft tumor experiment provided with furtherproof. Sensitivity to drug experiment has proved that CDD133+cells wereinsensitive to the clinical common chemotherapeutical drugs. These resultsdemonstrated they were capable of resistance to chemotherapy drugs andanti-apoptosis. Furthermore, ET-1stimulated the expression of vascularendothelial growth factor (VEGF) of CDD133+cells by regulation ofETAR, indicating the potential angiogenic properties of CD133+cells.Moreover, ET-1also reduced the degradation speed of HIF-1αby inhibitionof protease, therefore stabilized the expression of HIF-1α, which couldinduce the expression of vascular endothelial growth factor(VEGF) ofCD133+cells in hypoxia conditions.Conclusions:CD133+cells were successfully isolated from3AOovary cancer cell line and the biological properties of cancer stem cellswere characterized. ET-1contributes to the expression of vascularendothelial growth factor (VEGF) in CD133+cells and may influencethe angiogenic properties of CD133+cells. It also stabilizes the expressionof HIF-1α, which could enhance VEGF induction effect of CD133+cells inhypoxia conditions.