Panoply™ Human NT5E Knockdown Stable Cell Line

Recent studies have shown that the non-enzymatic functions of CD73 (also known as NT5E) play a key role in tumor progression, but its function in pancreatic cancer cells has not been deeply studied. In addition, the regulatory mechanism of CD73 in tumors is still unclear. Here, researchers found that CD73 is upregulated in pancreatic ductal adenocarcinoma (PDAC), and its expression is associated with poor prognosis. Knockdown of CD73 inhibits cell growth and induces G1 arrest through the AKT/ERK/cyclin D signaling pathway. Tumor necrosis factor receptor (TNFR) 2 is involved in the activation of AKT and ERK signaling pathways in CD73-induced PDAC. In addition, miR-30a-5p overexpression significantly enhanced the cytotoxic effect of gemcitabine in pancreatic cancer by directly targeting CD73 messenger RNA (mRNA), suggesting that the regulation of the miR-30a-5p/CD73 axis may play an important role in the formation of gemcitabine resistance in pancreatic cancer. In conclusion, this regulatory network of CD73 appears to represent a novel molecular mechanism for PDAC progression, and the interaction mechanism between miR-30a-5p, CD73, and TNFR2 may provide new insights into therapeutic strategies for pancreatic cancer.

In CD73 knockdown cells, cyclin D was significantly reduced, while the expression level of cyclin E was not changed (Figure 1a). In addition, in CD73 knockdown cells, the levels of G1 phase-related CDK4 and CDK6 did not change, while the level of p21 was slightly decreased (Figure 1a). To determine whether CD73 levels change throughout the cell cycle, the researchers examined the CD73 protein levels in PANC-1 cells synchronized by double thymidine blockade and harvested at different times (Figure 1b). The results showed that the expression of CD73 changed with the progression of the cell cycle and peaked at 4 hours and 11 hours, slightly earlier than cyclin D (Figure 1c). The AKT and MAPK signaling pathways have been shown to play an important role in regulating the cell cycle and can inhibit the expression of cyclin D, leading to G1 phase arrest. These data showed that the levels of p-AKT and p-ERK were significantly reduced in CD73 knockdown cells compared with control cells (Figure 1d). In contrast, the expression levels of p38 and JNK did not change. This suggests that CD73 knockdown may inhibit AKT/ERK activation, thereby reducing cyclin D expression and inhibiting the G1/S transition of pancreatic cancer cells.

Figure 1. CD73 knockdown induces G1 arrest via AKT/ERK/cyclin D signaling pathway. (Zhou L, et al., 2019)