Andrographolide induces protective autophagy and targeting DJ-1 triggers reactive oxygen species-induced cell death in pancreatic cancer

Zhaohong Wang; Hui Chen; Xufan Cai; Heqi Bu; Shengzhang Lin

doi:10.7717/peerj.17619

Andrographolide induces protective autophagy and targeting DJ-1 triggers reactive oxygen species-induced cell death in pancreatic cancer

PeerJ. 2024 Jun 28:12:e17619. doi: 10.7717/peerj.17619. eCollection 2024.

Authors

Zhaohong Wang^{1

2}, Hui Chen², Xufan Cai³, Heqi Bu⁴, Shengzhang Lin¹

Affiliations

¹ Department of Clinical Medicine, School of Medicine, Hangzhou City University, Hangzhou, China.
² Department of Hepatobiliary and Pancreatic Surgery, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
³ Zhejiang Chinese Medical University, Hanzhou, China.
⁴ Department of Surgery, Tongde Hospital of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou, China.

Abstract

Background: Andrographolide (Andro), an extract of Andrographis paniculate (Burm.f.) Wall. ex Nees (Acanthaceae), possesses diverse biologically active properties. However, the precise mechanisms and effects of Andro on pancreatic cancer (PC) remain unclear.

Methods: The cytotoxic potential of Andro and underlying mechanism towards PC cells was investigated through in vitro experiments and a xenograft mouse model. PC cells were first subjected to varying concentrations of Andro. The reactive oxygen species (ROS) was assessed using flow cytometry and DCFH-DA staining. The apoptosis rate was detected by flow cytometry. Additionally, western blot was applied to evaluate the expression levels of cleaved-caspase-3, DJ-1, LC3-I, LC3-II, and p62. To further elucidate the involvement of ROS accumulation and autophagy, we employed N-acetylcysteine as a scavenger of ROS and 3-Methyladenine as an inhibitor of autophagy.

Results: Andro demonstrated potent anti-proliferative effects on PC cells and induced apoptosis, both in vitro and in vivo. The cytotoxicity of Andro on PC cells was counteracted by DJ-1 overexpression. The reduction in DJ-1 expression caused by Andro led to ROS accumulation, subsequently inhibiting the growth of PC cells. Furthermore, Andro stimulated cytoprotective autophagy, thus weakening the antitumor effect. Pharmacological blockade of autophagy further enhanced the antitumor efficacy of Andro.

Conclusion: Our study indicated that ROS accumulation induced by the DJ-1 reduction played a key role in Andro-mediated PC cell inhibition. Furthermore, the protective autophagy induced by the Andro in PC cells is a mechanism that needs to be addressed in future studies.

Keywords: Andrographolide; DJ-1; Pancreatic cancer; Protective autophagy; Reactive oxygen species.

MeSH terms

Animals
Apoptosis* / drug effects
Autophagy* / drug effects
Cell Line, Tumor
Diterpenes* / pharmacology
Humans
Mice
Mice, Nude
Pancreatic Neoplasms* / drug therapy
Pancreatic Neoplasms* / metabolism
Pancreatic Neoplasms* / pathology
Protein Deglycase DJ-1* / genetics
Protein Deglycase DJ-1* / metabolism
Reactive Oxygen Species* / metabolism
Xenograft Model Antitumor Assays

Substances

andrographolide
Reactive Oxygen Species
Diterpenes
Protein Deglycase DJ-1
PARK7 protein, human

Grants and funding

This work was supported by the Natural Science Foundation of Zhejiang project (Y19H290009). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.