Fig. 3

Low concentrations of metformin reduced EMT of ovarian cancer cells and the CD44⁺CD117⁺ CSC population. SKOV3 cells were incubated with 0.1 mM metformin for 72 hours. The CD44⁺CD117⁺ cells were isolated by flow cytometry. Total RNA was isolated and mRNA was quantitated by real-time RT-PCR. mRNA expression of snail1, snail2, twist, vimentin, and E-cadherin was analyzed. Data show that treatment with 0.1 mM metformin reduced snail2, twist, and vimentin, but not snail1expression level, and upregulated the E-cadherin expression level (a). Bands detected with antibodies indicate the immune reactivity for snail1, snail2, twist, vimentin, E-cadherin, and β-actin: 72 hours of treatment with 0.1 nM metformin significantly downregulated snail2, twist, and vimentin, but not snail1, and upregulated E-cadherin as shown by western blotting (b). Total RNA was isolated from CD44⁺CD117⁺ cells and the mRNA was quantitated by real-time RT-PCR. Metformin at 0.1 mM significantly reduced snail2, twist, and vimentin, but not snail1 expression level, and upregulated the E-cadherin expression level (c). * Significant difference compared with control (P < 0.05)