Panoply™ Human EED Knockdown Stable Cell Line

Enhancer of zeste homolog 2 (EZH2) is activated in breast cancer, particularly triple-negative breast cancer (TNBC), and is crucial for cell invasion. It interacts with embryonic ectoderm development (EED) to maintain cancer stem cell (CSC) and epithelial-mesenchymal transition (EMT) characteristics, thereby promoting CSC metastasis. Since the binding of EZH2 to EED enhances EZH2's catalytic activity, inhibiting the EED-EZH2 interaction is a potential therapeutic strategy for EZH2-dependent cancers. Although several EED-EZH2 protein-protein interaction (PPI) inhibitors have been developed, few inhibitors specifically target EED. Here, researchers discovered that a genistein derivative compound (1) effectively binds to EED, thus blocking the EED-EZH2 PPI. Compound 1 was found to inhibit the proliferation of TNBC cells and suppress the growth of their three-dimensional tumor spheroids. Furthermore, by reversing EMT and reducing the proportion of CSCs, this compound inhibited the metastatic and invasive capabilities of TNBC. Therefore, targeting EED to disrupt the EED-EZH2 PPI may provide a novel approach for treating TNBC metastasis.

To verify that compound 1 reverses EMT and inhibits cancer stem cell biomarkers by targeting EED, researchers conducted overexpression experiments. Treatment with compound 1 significantly increased the protein levels of H3K27me3 and Snail in both EED-overexpressing and control MDA-MB-231 cells, and decreased the expression of E-cadherin and ALDH1A1. However, the biological effects of compound 1 were more pronounced in EED-overexpressing cells . Furthermore, the researchers constructed EED knockdown MDA-MB-231 cells (Figure 1A). Compared to control cells, EED knockdown cells showed lower levels of Snail and ALDH1A1, and higher expression levels of E-cadherin. Importantly, EED knockdown cells were more resistant to treatment with compound 1 in terms of the reduction of Snail, ALDH1A1, CD44, CD133, and H3K27me3, and the increase of E-cadherin, compared to control cells (Figures 1B-1H). In summary, these results indicate that compound 1 targets EED-EZH2, thereby inhibiting EMT and cancer stem cell biomarkers.

Figure 1. EZH2 is the direct target of compound 1. (Cheng S, et al., 2022)