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Published on: September 12, 2019
Artesunate Sensitizes human hepatocellular carcinoma to sorafenib via exacerbating AFAP1L2-SRC-FUNDC1 axis-dependent
Zhaochen Ma1, Wenjia Chen1, Yudong Liu1
1Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China.
Abstract:
Sorafenib is the most widely used first-line drug for the treatment of the advanced hepatocellular carcinoma (HCC). Unfortunately, sorafenib resistance often limits its therapeutic efficacy. To evaluate the efficacy of artesunate against sorafenib-resistant HCC and to investigate its underlying pharmacological mechanisms, a "sorafenib resistance related gene-ART candidate target" interaction network was constructed, and a signaling axis consisting with artesunate candidate target AFAP1L2 and sorafenib target SRC, and the downstream FUNDC1-dependent mitophagy was identified as a major contributor to the sorafenib resistance and a potential way of artesunate to mitigate resistance. Notably, our clinical data demonstrated that AFAP1L2 expression in HCC tissues was markedly higher than that in adjacent non-cancerous liver tissues (P < 0.05), and high AFAP1L2 expression was also significantly associated with an unfavorable overall survival of HCC patients (P < 0.05). Experimentally, AFAP1L2 was overexpressed in sorafenib resistant cells, leading to the activation of downstream SRC-FUNDC1 signaling axis, further blocking the FUNDC1 recruitment of LC3B to mitochondria and inhibiting the activation of mitophagy, based on both in vitro and in vivo systems. Moreover, artesunate significantly enhanced the inhibitory effects of sorafenib on resistant cells and tumors by inducing excessive mitophagy. Mechanically, artesunate reduced the expression of AFAP1L2 protein, suppressed the phosphorylation levels of SRC and FUNDC1 proteins, promoted the FUNDC1 recruitment of massive LC3B to mitochondria, and further overactivated the mitophagy and subsequent cell apoptosis of sorafenib resistant cells. In conclusion, artesunate may be a promising strategy to mitigate sorafenib resistance in HCC via exacerbating AFAP1L2-SRC-FUNDC1 axis-dependent mitophagy.Abbreviations: AFAP1L2, actin filament associated protein 1 like 2; ANOVA, analysis of variance; ANXA5, annexin V; ART: artesunate; CETSA, cellular thermal shift assay; CI: combination index; CO-IP: co-immunoprecipitation; CQ: chloroquine; CT, computed tomography; [18F]-FDG, fluoro-2-D-deoxyglucose F18; FUNDC1: FUN14 domain containing 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HCC, hepatocellular carcinoma; H&E Staining: hematoxylin - eosin staining; HepG2R, sorafenib resistant HepG2; IF, immunofluorescence; IHC, immunohistochemistry; LAMP1: lysosomal associated membrane protein 1; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; miR, microRNA; mRNA: messenger RNA; OE, overexpression; OS, overall survival; PET, positron emission tomography; qRT-PCR: quantitative real-time PCR; sh, short hairpin; shNC: negative control shRNA; shAFAP1L2: short hairpin AFAP1L2; SORA, sorafenib; SPR, surface plasmon resonance; SRC, SRC proto-oncogene, non-receptor tyrosine kinase; SUV, standardized uptake value; TEM, transmission electron microscopy; TOMM20: translocase of outer mitochondrial membrane 20.
Insights
Artesunate may overcome sorafenib resistance in hepatocellular carcinoma (HCC) by targeting the AFAP1L2-SRC-FUNDC1 pathway. This drug enhances mitophagy, promoting apoptosis in resistant HCC cells and tumors.
Area of Science:
- Molecular Biology
- Oncology
- Pharmacology
Background:
- Sorafenib is a first-line treatment for advanced hepatocellular carcinoma (HCC).
- Sorafenib resistance frequently limits its therapeutic effectiveness in HCC patients.
- Identifying novel therapeutic strategies to overcome sorafenib resistance is crucial.
Purpose of the Study:
- To evaluate the efficacy of artesunate (ART) against sorafenib-resistant HCC.
- To investigate the underlying pharmacological mechanisms of artesunate in overcoming resistance.
- To explore the role of the AFAP1L2-SRC-FUNDC1 signaling axis and mitophagy.
Main Methods:
- Construction of a gene-ART candidate target interaction network.
- In vitro and in vivo experiments using sorafenib-resistant HCC cell lines and tumor models.
- Analysis of AFAP1L2 expression, SRC and FUNDC1 phosphorylation, and mitophagy markers (LC3B).
Main Results:
- AFAP1L2 expression is upregulated in HCC tissues and associated with poor prognosis.
- AFAP1L2 overexpression activates the SRC-FUNDC1 axis, inhibiting mitophagy in resistant cells.
- Artesunate reduces AFAP1L2, suppresses SRC/FUNDC1 phosphorylation, and enhances mitophagy and apoptosis in resistant HCC.
Conclusions:
- Artesunate demonstrates efficacy against sorafenib-resistant HCC by targeting the AFAP1L2-SRC-FUNDC1 axis.
- Artesunate re-sensitizes resistant HCC cells to sorafenib by inducing excessive mitophagy.
- Artesunate represents a promising therapeutic strategy to overcome sorafenib resistance in HCC.
