Sanguinarine suppresses oral squamous cell carcinoma progression by targeting the PKM2/TFEB aix to inhibit autophagic flux

作者：Yong-Chun Peng, Zhijing He, Lun-cai Yin, Huifeng Pi, Yi Jiang, Ke-Yan Li, Li Tian, Jia Xie, Jian Zhang, Jian-Bo Zhang, Chenyao Li, Guan-Ying Feng, Kai Wang, Ding-Zhou Zhou, Xiaowei Xie, Zhiyuan Zhang, Tengfei Fan · 发表于：Phytomedicine · 年份：2024 · DOI：10.1016/j.phymed.2024.156337 · 被引用次数：24 · 研究领域：Autophagy in Disease and Therapy、Berberine and alkaloids research、Ginseng Biological Effects and Applications

BACKGROUND: Oral squamous cell carcinoma (OSCC) is one of the most common malignancies. However, there is no effective treatment for OSCC. PURPOSE: This study aimed to identify a natural compound with significant efficacy against OSCC and elucidate its primary mechanism of action. METHODS: An FDA-approved drug library and an MCE autophagy-related molecular compound library were screened through high-throughput screening to identify an effective natural compound against OSCC. The IC50 value of sanguinarine (Sang) in OSCC cells was determined using a CCK8 assay. Immunoblotting and immunofluorescence staining were used to assess the effect of Sang on autophagic flux in OSCC cells. Changes in the acidic lysosomal environment were evaluated using RFP-GFP-LC3B and LysoSensor Green DND-189. Furthermore, limited proteolysis-coupled mass spectrometry (LiP-MS) and virtual screening techniques were utilized to identify direct binding targets of Sang, which were subsequently validated by surface plasmon resonance (SPR) and microscale thermophoresis (MST). Molecular docking combined with molecular dynamics analysis identified the binding site between the target protein and Sang. In vitro and in vivo investigations with mutant plasmids confirmed this finding. RESULTS: Screening led to the identification of the naturally occurring autophagy modulator Sang as a potent inhibitor of OSCC progression. Moreover, Sang impaired lysosomal function through reducing lysosomal-associated membrane prot...