2A, the proliferation of SKOV-3

was inhibited significantly on 3rd d while there was no difference after 7d’s incubation. As for the proliferation of ES-2 cells, there has no significant difference after incubation under hypoxia. The proliferation of HUVEC cells were inhibited by incubation under hypoxia for 3 d and further inhibited after 7 d’s incubation. Figure 2 The proliferation, cell cycle, apoptosis, Quizartinib order invasion of SKOV-3, ES-2 and HUVEC cells induced by hypoxia. The SKOV-3, ES-2 and HUVEC cells were cultured for 3 or 7 d in normoxia or hypoxia conditions before proliferation, cell cycle (S-phage), apopotosis and invasion detected by MTT, FCM (for cell cycle and apoptosis) and Transwell as shown in methods. GW786034 in vivo A. The proliferation of three cells by MTT. B. The S-phase ratio in three cells by FCM. C. The apoptosis of three cells detected by FCM. D and E. The numbers of cells invasion through the membrane indicated by Transwell after incubated for 3 days (D) or 7 days (E). Data were shown in Mean ± S.D. from three separate experiments with the similar result. * and ** indicates P < 0.05 and P < 0.01 vs. Normoxia. The percent of cells in S-phase and apoptosis after incubation for 3 or 7 d under hypoxia were shown in Fig. 2B and 2C. As they shown, in the case of SKOV-3

SHP099 and ES-2 cells, the percent in S-phase were decreased and those of apoptosis were increased after 3 d’s incubation, however, there had no difference in S-phase and apoptosis after 7 d’s incubation of the two cell lines. On the other hand, the percent of S-phase of HUVEC cells was decreased and that of apoptosis was increased after both 3 and 7 d’s incubation. The numbers of cell migrated through basement membrane of the transwell chamber were shown in Fig. 3D (after 3 d’s incubation) and 3E (after 7 d’s incubation). Compared to normoxia control, the numbers decreased significantly in SKOV-3 after 3 and 7 d’s incubation under hypoxia while it decreased significantly in ES-2 only after 3 d’s incubation. The numbers of HUVEC cells were decreased significantly after both

3 and 7 d’s incubation. Figure 3 The genes expression in SKOV-3, ES-2, ELs from cancer cells and HUVEC induced by hypoxia. The SKOV-3, ES-2 and Plasmin HUVEC cells were cultured for 7 d in normoxia or hypoxia conditions before harvested for the expression of HIF-1a, VEGF, Flk-1, CyclinD1, p53 and V-src genes detected by Real-time PCR. A. The genes expression in SKOV-3 and relative cells by Real-time PCR. B. The genes expression in ES-2 and relative cells by Real-time PCR. SKOV-3 EL: the endothelial-like cells induced from SKOV-3 cells; SKOV-3+Si: the SKOV-3 cells treated by Sirolimus under hypoxia; ES-2 EL: the endothelial-like cells induced from ES-2 cells; ES-2+Si: the ES-2 cells treated by Sirolimus under hypoxia; *, ^, and & indicates that P < 0.05 vs.HUVEC, SKOV-3 (or ES-2) and SKOV-3+Si (or ES-2+Si); **, ^^, and && indicates that P < 0.01 vs.HUVEC, SKOV-3 (or ES-2) and SKOV-3+Si (or ES-2+Si).