Figure 6
EGCG regulated the interaction between JWA and topoisomerase IIα and their synergistic effect on inhibition of NCI-H460 cells migration and invasion.
(a) NCI-H460 cells were transfected with siRNA-topoisomerase IIα (100 pmol), shRNA-JWA, JWA and topoisomerase IIα plasmids (4 μg) as well as the control vector. Migration ability of the cells at various time points after transfection (24 h, 48 h) was assessed by scratch migration assay. (b) The JWA or topoisomerase IIα plasmid (4 μg) was transiently transfected into NCI-H460 cells. 24 hours later, target proteins in cell lysates were detected by immunoblotting using antibodies against MMP-2/9, N- cadherin, ZEB1, slug, snail and E-cadherin. β-actin expression served as a loading control. (c) and (d) NCI-H460 cells were transfected with Flag-JWA , Flag- topoisomerase IIα and Flag-vector plasmids, (4 μg) in the presence or absence of EGCG (40 μM) for 24 h. The cells were harvested and lysed for the detection expression of JWA or topoisomerase by Western blot analysis. β-actin expression served as a loading control. Error bars represent the mean ± SD of triplicate experiments. Statistical differences to the controls were shown as *p < 0.05, **p < 0.01.
