Panoply™ Human NUAK1 Over-expressing Stable Cell Line

Metastasis remains a major cause of poor pathological outcomes and prognosis in patients with esophageal squamous cell carcinoma (ESCC). NUAK1 has been reported to be highly expressed in various human cancers and is associated with poor prognosis in cancer patients. However, the role of NUAK1 in ESCC metastasis and its underlying signaling pathway mechanisms remain unclear. Here, the study shows that NUAK1 is highly expressed in ESCC tissues compared to adjacent normal esophageal epithelial tissue. Furthermore, high NUAK1 expression is positively correlated with tumor invasion depth, lymph node metastasis, pathological TNM stage, and survival in ESCC patients. Further experiments showed that NUAK1 overexpression does not affect the viability and clonogenic capacity of ESCC cells, but significantly promotes cell migration and invasion in vitro and experimental lung metastasis in vivo. Mechanistically, NUAK1 enhances the transcriptional level of Slug, thereby enhancing the migration and invasion capabilities of ESCC cells. Sustained silencing of the Slug gene almost completely inhibits the migration and invasion capabilities of NUAK1-overexpressing ESCC cells. Further research showed that NUAK1 upregulates the transcriptional activity of Slug by activating the JNK/c-Jun signaling pathway. Therefore, NUAK1 promotes the metastasis of ESCC cells by activating the JNK/c-Jun/Slug signaling pathway, suggesting that NUAK1 is a potential therapeutic target for metastatic ESCC.

Here, researchers sought to determine the potential role of NUAK1 in the migration and invasion of ESCC cells. Scratch healing assays showed that NUAK1-overexpressing EC109 and KYSE510 cells exhibited significantly enhanced wound healing capabilities (Figure 1A). Similarly, Transwell migration assays also demonstrated significantly enhanced migration ability in NUAK1-overexpressing ESCC cells (Figure 1B). Transwell invasion assays showed that NUAK1 overexpression significantly increased the number of invading ESCC cells (Figure 1C). Given that NUAK1 overexpression promotes ESCC cell migration and invasion, the researchers further investigated whether silencing NUAK1 could attenuate these malignant behaviors. As shown in Figure 1D, knockdown of NUAK1 significantly inhibited the migration ability of KYSE30 and KYSE150 cells. Consistent with this result, Transwell migration assays also showed that NUAK1-knockdown ESCC cells exhibited significantly reduced migration ability compared to shNC-transfected cells (Figure 1E). Furthermore, the number of tumor cells crossing Matrigel was reduced in both types of NUAK1 knockdown ESCC cells (Figure 1F). Therefore, these data suggest that NUAK1 overexpression promotes the migration and invasion of ESCC cells.

Figure 1. NUAK1 promotes the migration and invasion of ESCC cells. (Yang H, et al., 2023)