2. Academic Validation
  3. Inhibitory Effects of Esculetin on Liver Cancer Through Triggering NCOA4 Pathway-Mediation Ferritinophagy in vivo and in vitro

Inhibitory Effects of Esculetin on Liver Cancer Through Triggering NCOA4 Pathway-Mediation Ferritinophagy in vivo and in vitro

  • J Hepatocell Carcinoma. 2023 Apr 11;10:611-629. doi: 10.2147/JHC.S395617.
Zhiru Xiu 1,  Yiquan Li 1,  Jinbo Fang 1,  Jicheng Han 1,  Shanzhi Li 1,  Yaru Li 1 2,  Xia Yang 1,  Gaojie Song 3,  Yue Li 1,  Ningyi Jin 1 4 5,  Yilong Zhu 1,  Guangze Zhu 1,  Lili Sun 6,  Xiao Li 1 4 7
Affiliations

Affiliations

  • 1 Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun, People's Republic of China.
  • 2 Medical College, Yanbian University, Yanji, People's Republic of China.
  • 3 Medical College, Jiujiang· University, Jiujiang, People's Republic of China.
  • 4 Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, People's Republic of China.
  • 5 Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, People's Republic of China.
  • 6 Department of Head and Neck Surgery, Tumor Hospital of Jilin Province, Changchun, People's Republic of China.
  • 7 College of Life Sciences, Shandong Normal University, Jinan, People's Republic of China.
PMID: 37069958 DOI: 10.2147/JHC.S395617
Abstract

Objective: To explore the effects of Esculetin on liver Cancer and explore potential mechanisms of Esculetin-inducing cells death.

Methods: Esculetin's effects on the proliferation, migration and Apoptosis of HUH7 and HCCLM3 cells were detected by using CCK8, crystal violet staining, wound healing, TranswellTM and Annexin V-FITC/PI. Flow cytometry, fluorescence staining, Western blot, T-AOC, DPPH radical scavenging assay, hydroxyl radical's inhibitory capability and GSH test were used to examine the esculetin's effects on the ROS level, the oxidation-related substances and proteins' expression in hepatoma cells. In vivo experiment was performed by xenograft model. Ferrostatin-1 was used to determine the death way of hepatoma cells induced by esculetin. Live cell probe, Western blot, Fe2+ content, MDA, HE staining, Prussian blue staining and immunohistochemistry were used to examine the ferritinophagy-related phenomenon induced by esculetin in hepatoma cells. The relationship between esculetin and NCOA4-mediated ferritinophagy was confirmed through gene silence and overexpression, immunofluorescence staining and Western blot.

Results: Esculetin suppressed the proliferation, migration and Apoptosis of HUH7 and HCCLM3 cells significantly, influenced the oxidative stress level, altered the Autophagy and iron metabolism levels in cells, and produced a ferritinophagy-related phenomena. Esculetin increased the levels of cellular lipid peroxidation and Reactive Oxygen Species. In vivo, esculetin could decrease tumour volume, promote LC3 and NCOA4 expressions, suppresse hydroxyl radical's inhibiting capacity and GSH, increase Fe2+ and MDA levels, decrease antioxidant proteins expression in tumour tissue. In addition, Esculetin could also increase the iron deposition of tumour tissues, promote ferritinophagy, and induce tumours' Ferroptosis.

Conclusion: Esculetin has an inhibitory effect on liver Cancer in vivo and in vitro through triggering NCOA4 pathway-mediation ferritinophagy.

Keywords

NCOA4; anti-tumour; esculetin; ferritinophagy; liver cancer.

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