Twist promotes epithelial-mesenchymal transition (EMT) markers.

<p>(A) Expression of Twist, ERα, E-cadherin and Vimentin in breast cancer cell lines was analyzed by Western blotting. β-actin was used as a loading control. (B) The transcriptional level of Twist, ERα, E-cadherin and Vimentin were detected by Real-Time PCR analysis. All the data were collected from three independent experiments. (C and E) Overexpression of Twist resulted in a loss of E-cadherin and a gain of Vimentin and Snail in MCF-7 and T47D cells. The expression of ERα, E-cadherin, Vimentin and Snail was detected by western blot analysis. β-actin was used as the loading control. (D and F) Overexpression of Twist resulted in a decrease of E-cadherin and an increase of Vimentin and Snail in mRNA level in MCF-7 and T47D cells. The mRNA expression levels for each target gene were subsequently normalized by the mRNA level of β-actin. All samples were measured in triplicate and <i>P</i> value low than 0.05 for MCF-7 and T47D cells overexpressing Twist compared with the corresponding MCF-7 and T47D cells transfected with empty vector. Data shown are means ± SD of at least three independent experiments. <i>P</i>-values were obtained using the Student’s <i>t</i>-test analysis. Western blot was used to detected Twist and EMT marker expression (n = 3).