Panoply™ Human MAP3K8 Knockdown Stable Cell Line

Superenhancers (SEs) regulate gene expression, which is crucial for cell type identification and tumorigenesis. Here, researchers used chromatin immunoprecipitation sequencing (ChIP-seq) and high-resolution chromatin immunoprecipitation sequencing (HiChIP-seq) to discover that mitogen-activated protein kinase 8 (MAP3K8) is an SE-related gene that interacts with chromosomes in Epstein-Barr virus-associated gastric cancer (EBVaGC) cells. CRISPRi-mediated inhibition of MAP3K8 SEs attenuated MAP3K8 expression and EBVaGC cell proliferation. Furthermore, functional analysis of MAP3K8 in EBVaGC cells showed that silencing MAP3K8 inhibited EBVaGC cell proliferation, colony formation, and migration. RNA sequencing and pathway analysis indicated that knockdown of MAP3K8 blocked the Notch signaling pathway and epithelial-mesenchymal transition (EMT) in EBVaGC cells. Analysis of the Cancer Genome Atlas (TCGA) and GSE51575 databases showed that MAP3K8 expression was enhanced in gastric cancer and negatively correlated with disease-free survival. Furthermore, Spearman correlation analysis showed that MAP3K8 expression was positively correlated with the expression of Notch pathway and EMT-related genes (such as Notch1, Notch2, C-terminal binding protein 2 (CTBP2), α-smooth muscle actin isoform 2 (ACTA2), transforming growth factor β receptor 1 (TGFβR1), and Snail family transcriptional repressor 1/2 (SNAI1/SNAI2)) in gastric cancer.

Here, researchers constructed MAP3K8 knockdown SNU719 and YCCEL1 cell lines. The study showed that cell proliferation was reduced in MAP3K8 knockdown cells (Figures 1A-D). Furthermore, inhibiting MAP3K8 limited the colony-forming ability of SNU719 and YCCEL1 cells (Figures 1E-H). Boyden chamber-assisted cell migration assays demonstrated that the migration ability of EBVaGC cells was reduced in MAP3K8 knockdown cells (Figures 1I-L).

Figure 1. Functional analysis of MAP3K8 in EBVaGC. (Roy G, et al., 2023)