**2. Methods**

### **2.1 Ovarian cancer cell biological behaviors affected by ivermectin**

The normal ovarian cells IOSE80 and ovarian cancer cells TOV-21 and SKOV3 were treated with ivermectin to measure ivermectin-mediated ovarian cancer cell biological behavior changes. (i) IOSE80, TOV-21G, and SKOV3 were treated with ivermectin (0–60 μM) for 24 h, followed by the use of CCK8 to measure the IC50 of ivermectin in each cell. (ii) TOV-21G and SKOV3 were treated with ivermectin (0 μM, 10 μM, 20 μM, and 30 μM) for 24 h, followed by the use of EdU assay to measure DNA synthesis in each cell. (iii) TOV-21G and SKOV3 were treated with ivermectin (0 μM, 10 μM, 20 μM, and 30 μM) for 48 h, followed by clonogenic

*The Anti-Cancer Effects of Anti-Parasite Drug Ivermectin in Ovarian Cancer DOI: http://dx.doi.org/10.5772/intechopen.95556*

assay to measure the *in vitro* effects of ivermectin in each cell. (iv)TOV-21G and SKOV3 were treated with ivermectin (0 μM, 10 μM, 20 μM, and 30 μM) for 24 h, followed by flow cytometry to measure cell cycle and cell apoptosis changes in each cell. (v) When A2780 and TOV-21G seeded in 6-well plates were grown to approximately 90% confluency, followed by the use of 10-μl pipette tip to make an artificial wound, and then treated with ivermectin (0 μM, 10 μM, 20 μM, and 30 μM) for 24 h, and measure the wound healing. The relative percentage of wound healing = (the width of wound at 0 h the width of wound at 24 h)/the width of wound at 0 h. The detailed procedure was described previously [4, 21].
