Fig. 5: miR-221-3p is regulated directly by TWIST2

a Schematic structure of the miR-221-3p upstream promoter containing a TWIST2-binding site. b Luciferase activity of PGL3-221-3p construct after transfection of TWIST2 plasmid in SiHa and 293 T cells. c Western blotting analysis of expression of E-cadherin, N-cadherin, and Vimentin proteins in SiHa, SiHa-tw2, SiHa-shwt2, or SiHa-shtw2 cells transfected with miR-221-3p mimic or miR-221-3p inhibitor; and HeLa cells transfected with TWIST2 expression vector with/without miR-221-3p mimic or TWIST2 siRNA with/without miR-221-3p inhibitor. β-actin was used as loading control. d Cell migration was quantified as the percentage of wound-healed area; average number of invaded cells per field from three independent experiments. Data represent means ± SD of five randomly selected areas (*p < 0.05; **p < 0.001). e IHC analysis of TWIST2 and in situ hybridization analysis of miR-221-3p in matched cervical specimens. Human CC tumors with high TWIST2 expression and had strong miR-221-3p expression. Normal cervical tissues with low TWIST2 expression and had weak miR-221-3p expression. Representative images are shown at ×400 magnification. The HSCOREs of miR-221-3p and TWIST2 in different cervical specimens are also shown (*p < 0.05) (upper-right panel). Correlation of miR-221-3p staining and TWIST2 staining was analyzed (lower-right panel)