Panoply™ Human AURKA Knockdown Stable Cell Line

Aurora kinase A (AURKA) is involved in cell cycle progression, mitosis, and the regulation of a series of key oncogenic signaling pathways in various malignancies, including neuroblastoma. Small molecule inhibitors of AURKA have shown some potential, but their efficacy in clinical trials has not yet met expectations. Here, researchers evaluated the inhibitory effect of the AURKA inhibitor MLN8237 on neuroblastoma cells to understand its potential mechanisms of action in cancer therapy. The researchers found that the AURKA inhibitor MLN8237 induced cellular senescence and G2/M cell cycle arrest in the neuroblastoma cell line IMR32. AURKA inactivation led to MYCN instability and inhibited cell growth in vitro and in vivo mouse models. Although MLN8237 inhibited AURKA kinase activity, it had little inhibitory effect on AURKA protein levels. Conversely, MLN8237 treatment resulted in abnormally high expression of AURKA in vitro and in vivo. Knockdown of AURKA reduced cell viability. The combination of MLN8237 and AURKA small interfering RNA showed a more significant inhibitory effect on neuroblastoma cell growth. Furthermore, the combination of MLN8237 treatment and AURKA siRNA induced apoptosis in senescent cells by inhibiting the Akt/Stat3 pathway.

Since AURKA is abnormally highly expressed in neuroblastoma cells treated with MLN8237, knocking down AURKA should have an inhibitory effect on cell growth. Here, researchers constructed AURKA knockdown IMR32 cells (Figure 1a). The results showed that knocking down AURKA induced apoptosis and inhibited cell growth (Figure 1b, c). No effect on cell cycle arrest was observed in AURKA knockdown cells (Figure 1d). SA-β-gal staining showed no senescent cells in AURKA knockdown cells. Since AURKA can activate the phosphorylation and transcriptional activity of Stat3, the researchers evaluated the Jak2/Stat3 pathway and found that knocking down AURKA reduced the activity and protein levels of both Jak2 and Stat3 (Figure 1e). Although the protein levels of Akt were not affected in AURKA knockdown cells, phosphorylated Akt was significantly reduced (Figure 1e). Therefore, AURKA knockdown induces apoptosis and reduces cell survival by inhibiting the Jak2/Stat3 pathway and inactivating Akt.

Figure 1. AURKA knockdown induced cell apoptosis and cell growth inhibition by repressing the activity of Akt/Stat3 pathway. (Yang Y, et al., 2020)