RNA-Based TWIST1 Inhibition via Dendrimer Complex to Reduce Breast Cancer Cell Metastasis

Figure 4

TWIST1 knockdown following YTZ3-15 delivery of siTwist decreases cell motility and downstream EMT marker expression. (a) Compared to siQ control (at seven days), siTwistA (TwA) and siTwistB (TwB) delivered via YTZ3-15 produced >90% TWIST1 knockdown at the mRNA level. Knockdown lasted seven days after transfection. (b) Compared to siQ control (at seven days), TwA and TwB delivered via YTZ3-15 produced knockdown of the TWIST1 targets N-Cadherin and Vimentin. N-Cadherin mRNA levels decreased by >40% after one day, and by approximately 90% after seven days. Vimentin mRNA was nearly undetectable after one day, and remained at <10% after seven days. (c) YTZ3-15 transfection of siTwistA decreased directional migration compared to siQ transfected cells (control) in wound healing assays. Dashed lines indicate migratory front and were placed manually. Images shown are representative data from experiments performed in triplicate. (d) Left: YTZ3-15 transfection of TwA or TwB resulted in >50% decrease in invasion of SUM 1315 cells through Matrigel. Cells were allowed to migrate for one day, following one day incubation with YTZ3-15-siRNA dendriplexes. Five fields per condition were imaged (representative images shown). Right: quantification of image data. Bars represent mean and standard deviation of five fields per condition.

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