Hernandezine induces autophagic cell death in human pancreatic cancer cells via activation of the ROS/AMPK signaling pathway

Chang-feng Song1,2,3,4, Yu-heng Hu1,2,3,4, Zhi-guo Mang5, Zeng Ye1,2,3,4, Hai-di Chen1,2,3,4, De-sheng Jing1,2,3,4, Gui-xiong Fan1,2,3,4, Shun-rong Ji1,2,3,4, Xian-jun Yu1,2,3,4, Xiao-wu Xu1,2,3,4, Yi Qin1,2,3,4
1 Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, China
2 Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
3 Shanghai Pancreatic Cancer Institute, Shanghai 200032, China
4 Pancreatic Cancer Institute, Fudan University, Shanghai 200032, China
5 Yujie Biotechnology (Shanghai) Co., Ltd., Shanghai 201314, China
Correspondence to: Xian-jun Yu:, Xiao-wu Xu:, Yi Qin:,
DOI: 10.1038/s41401-022-01006-1
Received: 28 November 2021
Accepted: 14 March 2022
Advance online: 25 October 2022


Hernandezine (Her) is a bisbenzylisoquinoline alkaloid extracted from the traditional Chinese herbal medicine Thalictrum glandulosissimum. Evidence shows that Her is a natural agonist of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and induces apoptosis and autophagy in tumor cells. In this study, we investigated the role of autophagy in Her-induced cell death in human pancreatic cancer cell lines. We showed that Her dose-dependently suppressed cell proliferation, promoted autophagy and induced autophagic death in pancreatic ductal adenocarcinoma (PDAC) cell lines Capan-1 and SW1990. The IC50 values of Her in inhibition of Capan-1 and SW1990 cells were 47.7 μM and 40.1 μM, respectively. Immunoblotting showed that Her (1−40 μM) promoted the conversion of LC3-I to LC3-II, and Her exerted concentration-dependent and time-dependent effects on autophagy activation in PDAC cells. In transmission electron microscopy and fluorescence image analysis, we found that autophagic vacuoles were significantly increased in Her-treated cells. Knockdown of ATG5, a key gene in the autophagy pathway, alleviated the activation of autophagy by Her. These results demonstrated that Her induced autophagy in PDAC cells. Intensely activated autophagy could promote cell death. The autophagy inhibitors, BafA1 and HCQ significantly inhibited Her-induced cell death, implying that Her induced autophagic cell death in PDAC cells. Moreover, we showed that Her activated autophagy by increasing the phosphorylation of AMPK and decreasing the phosphorylation of mTOR/p70S6K. Knockdown of AMPKα relieves the autophagic cell death induced by Her. Furthermore, Her concentration-dependently enhanced reactive oxygen species (ROS) generation in PDAC cells. Antioxidants could reduce the phosphorylation of AMPK and suppress autophagic cell death induced by Her. Our study provides evidence for the development of Her as a therapeutic agent for the treatment of pancreatic cancer.
Keywords: pancreatic cancer; hernandezine; ROS; autophagy; autophagic cell death

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