Chelerythrine (CHE) significantly reduced tumor size and weight in nude mice bearing KYSE150 tumors.
CHE inhibited the proliferation, migration, and invasion of esophageal squamous cell carcinoma (ESCC) cells in a dose-dependent manner.
The compound induced cell apoptosis and triggered cell death through a specific pathway involving PINK1 and Parkin, linked to increased reactive oxygen species in mitochondria.
CHE altered the production of autophagosomes and autolysosomes when used with autophagy inhibitors, indicating its role in complete autophagic flux.
The treatment affected various signaling pathways related to protein degradation, , and mitochondrial energy metabolism.
CHE also reduced the presence of M2 macrophages in the tumor microenvironment and influenced cell metabolism.
Simplified
BACKGROUND: Chelerythrine (CHE) exhibits notable anti-inflammatory and antitumor properties, while its impact on esophageal squamous cell carcinoma (ESCC), especially the underlying mechanisms remain unclear. In this study, we aim to investigate the roles and mechanism of CHE in the ESCC treatment.
METHODS: Human ESCC cell lines and organoids were used for in vitro cell experiments, BALB/c nude mice were used for in vivo animal experiments. To investigate the underlying mechanism of CHE treatment, drug library screen, RNA sequencing analysis, TMT-based quantitative proteomic analysis, western blotting analysis, immunofluorescence, immunohistochemistry, quantitative real-time polymerase chain reaction, mitochondrial membrane potential assay, apoptosis assay, detection of mitochondrial reactive oxygen species (mtROS), autophagic flux monitoring, transmission electron microscopy, and seahorse XF-96 metabolic flux analysis were used to assess the effect of CHE and relevant mechanism.
RESULTS: CHE dose-dependently inhibited the proliferation, migration, and invasion of ESCC cells. CHE also induced cell apoptosis and triggered PTEN-induced kinase 1 (PINK1)-Parkin-mediated -mediated cell death by elevating the production of reactive oxygen species in mitochondria and diminishing mitochondrial membrane potential (MMP). However, the production of autophagosomes and autolysosomes induced by CHE altered when used in combination with the autophagy inhibitors 3-methyladenine (3-MA) or bafilomycin A1 (BafA1), indicating that it induced complete autophagic flux in the cells. Mechanistically, CHE affected multiple signaling pathways associated with ubiquitin-mediated proteolysis, mitophagy, and mitochondrial energy metabolism, indicating its close involvement in mitophagy occurrence. In addition, CHE treatment significantly reduced tumor size and weight in nude mice bearing KYSE150 tumors and retarded the growth of organoids derived from patients, it also reduced the ratio of M2 macrophage in tumor microenvironment and cell metabolism.
CONCLUSIONS: CHE activates PINK1-Parkin-mediated mitophagy and disrupts mitochondrial homeostasis, and it also affects the tumor environment and cell metabolism, ultimately leading to cell death, supporting the potential of CHE for ESCC therapy.
Key numbers
> 90%
Inhibition Rate
Percentage inhibition of cell viability at 30 µM concentration.
5.616 µM, 10.80 µM, 10.76 µM
Values
Half maximal inhibitory concentration for KYSE30, KYSE150, and KYSE450 cells, respectively.
Significantly decreased
Tumor Weight Reduction
Tumor weight in -treated mice vs. control group.
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Declarations. Ethics approval and consent to participate: All mouse experiments were performed in compliance with the ethical policies and procedures approved by the Institutional Animal Care and Use Committee of Xiamen University (Approval no. XUMLAC20200202). Animal experiment procedures were performed according to the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and followed the guidelines of the Animal Welfare Act. Consent for publication: All authors have read and approved the final manuscript. Competing interests: The authors declare that they have no competing interests.
