Triptolide enhances TRAIL sensitivity of pancreatic cancer cells by activating autophagy via downregulation of PUM1.

BACKGROUND: Triptolide (TPL) can enhance the sensitivity of pancreatic cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), but available research is limited to whether TPL can affect the relevant downstream signaling pathways of TRAIL. Current knowledge is far from adequate to fully understand the mechanisms by which TPL increases TRAIL sensitivity of pancreatic cancer. PURPOSE: We aimed to find TPL-regulated upstream components of the signaling pathways of TRAIL to further understand the regulatory mechanism by which TPL increases the sensitivity to TRAIL. METHODS: Microarray analysis and the adherent cell cytometry system Celigo were used to identify the TRAIL-related genes. Western blot analysis, cell proliferation assays, tumorigenicity assays in nude mice, flow cytometry, and transmission electron microscopy were performed to analyze the function of Pumilio RNA-binding family member 1 (PUM1) in TPL-mediated enhancement of sensitivity to TRAIL. The effect of PUM1 silencing on the p27-CDK2 complex was examined by immunoprecipitation. RESULTS: PUM1 expression was decreased by TPL and TPL + TRAIL but was not decreased by TRAIL alone. PUM1 silencing enhanced low-concentration-TRAIL-induced suppression of proliferation and promotion of apoptosis and increased p27 expression and the amount of the p27-CDK2 complex in pancreatic cancer cells. PUM1 overexpression attenuated the effects of TPL treatment (TRAIL-induced cell proliferation suppression and apoptosis promotion), while PUM1 silencing and TPL enhanced low-concentration-TRAIL-induced autophagy activation in pancreatic cancer cells. Moreover, PUM1 overexpression attenuated the effect of TPL treatment on TRAIL-induced autophagy activation in pancreatic cancer cells. CONCLUSION: PUM1 silencing increased the sensitivity of pancreatic cancer cells to TRAIL in vivo and in vitro, indicating that PUM1 may be a new target for increasing the sensitivity of cancer cells to TRAIL. In addition, our results indicate that TPL enhances TRAIL sensitivity of pancreatic cancer cells by activating autophagy via downregulation of PUM1. This novel concept may have significant implications for the development of new strategies to enhance TRAIL sensitivity of tumors.