Antitumor effect As shown in Figure 2-A, the viability of cells dose-dependently reduced. GCV at the density of 10-2-103 μg/ml had obvious antitumor effect on SKOV3/tk (IC50:2.24 ± 0.23 μg/ml) and SKOV3/tk-MCP-1 (IC50:2.06 ± 0.31 μg/ml). The IC50 value of SKOV3/tk and SKOV3/tk-MCP-1 significantly dropped when compared to that of SKOV3/neo (P < 0.05). There was no significant

difference between SKOV3/MCP-1 group and control groups (P > 0.05). Besides, the beginning cytotoxic time of 0.1 μg/ml GCV and 1.0 μg/ml GCV was both 48 h, and the 96 h kill rate of 0.1 μg/ml GCV and 1.0 μg/ml GCV against SKOV3/tk-MCP-1 was 40 ± 2.19% and 90 ± 4.55% respectively (P < 0.05) (Figure 2-B). Figure 2 Antitumor effection. A: MTT assay of GCV on ovarian cancer cells. B: GCV at check details the density of 0.1 μg/ml, the beginning cytotoxic was 48 h and 40% kill rate at 96 h, however, the beginning cytotoxic was 48 h and AZD9291 90% kill rate at 96 h when GCV at the density of 1.0 μg/ml.

C: Lethal effect of mononuclear macrophage on SKOV3/MCP-1 and SKOV3/tk-MCP-1 was determined by MTT assay. D: There is a synergistic antitumor effect when cooperated tk-MCP-1 + GCV system with mononuclear macrophage. The antitumor effect of monocytes on ovarian cancer cells: The maximum lethality rate of SKOV3/MCP-1 and SKOV3/tk-MCP-1 was 29 ± 1.25% and 23 ± 2.18% respectively, comparing to 1.8 ± 0.64% of SKOV3/neo (P < 0.05). We found that the lethal effect of monocytes on tumor cells was effector-dependent, and the maximum lethality rate appeared at the ratio of 20:1(Figure GNA12 2-C). The survival rate of SKOV3/tk and SKOV3/tk-MCP-1 incubating with SKOV3 in different ratio was

evaluated after addition GCV or GCV plus monocytes (Figure 2-D). When 10 μg/ml GCV was added, only 10% of SKOV3/tk or SKOV3/tk-MCP-1 could kill about 40% of tumor cells. When the ratio of SKOV3/tk or SKOV3/tk-MCP-1 to SKOV3 was 50%, there were about 80% of tumor cells killed. But cytotoxin did not appear with SKOV3/neo(P < 0.05). Only 10% of tk-MCP-1 + GCV + monocytes system could kill about 70% of tumor cells, while 40% of tk-MCP-1 + GCV + monocytes system could kill about 90% of tumor cells. The result of flow cytometer showed that the apoptotic rate of SKOV3/tk-MCP-1 (13.48 ± 1.01%) was obviously higher than those of SKOV3/tk (9.50 ± 1.33%) and SKOV3/neo (2.19 ± 0.56%) (P < 0.05), S phase of SKOV3/tk (38.31 ± 1.67%) was lower than that of SKOV3/tk-MCP-1 (52.92 ± 1.78%) (P < 0.05)(Table 1). Table 1 Post-treatment apoptotic rate and cell cycle analysis ( )   SKOV3/neo SKOV3/tk SKOV3/tk-MCP-1 Apoptotic rate (%) 2.19 ± 0.56 9.50 ± 1.33 13.48 ± 1.01 G0/G1 (%) 53.90 ± 1.66 53.10 ± 1.21 40.28 ± 1.11 S (%) 19.34 ± 0.65 38.31 ± 1.67 52.92 ± 1.78 G2/M (%) 26.76 ± 1.01 8.59 ± 1.25 6.80 ± 1.