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Osthole inhibits GSK-3β/AMPK/mTOR pathway-controlled glycolysis and increases radiosensitivity of subcutaneous transplanted hepatocellular carcinoma in nude mice

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Abstract

Purpose

Osthole possesses anti-tumor activities. However, whether osthole can have a radiosensitization effect on hepatic cancer remains unclear. Here, an HCC-LM3 cells-inoculated subcutaneous transplanted tumor was adopted to explore the effect of osthole.

Methods

The tumor-bearing mice were treated with 100 mg/kg osthole for 12 days, 4 Gy irradiation twice, or their combination. The tumor volume and weight, lactic acid content, glycolytic enzyme activities, and protein expression of glycogen synthase kinase 3β (GSK-3β), p‑GSK-3β, mammalian target of rapamycin (mTOR), p‑mTOR, AMP-activated protein kinase (AMPK), p‑AMPK, glucose transporter 1/3, and pyruvate kinase M2 were determined. The GSK-3β-overexpressed HCC-LM3 or SK-Hep‑1 cell models were also adopted to verify the effects of osthole on expression of these proteins.

Results

The tumor volume and weight, lactic acid content, and glycolytic enzyme activities in tumor tissues were lower in the osthole + radiation group than in the radiation group. Moreover, osthole could reverse the radiation-induced increments of p‑GSK-3β/GSK-3β and p‑mTOR/mTOR protein ratios and the expression of glucose transporter 1/3 and pyruvate kinase M2 proteins in tumor tissues, and increase the protein ratio of p‑AMPK/AMPK. The effects of osthole on these glycolysis-related proteins were also observed in GSK-3β-overexpressed HCC-LM3 or SK-Hep‑1 cell models.

Conclusion

Osthole has a radiosensitizing effect on subcutaneous transplanted hepatocellular carcinoma, and its mechanism may be related to inhibition of GSK-3β/AMPK/mTOR pathway-controlled glycolysis.

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Funding

This work was supported by the Research Fund of Nanjing Medical University (NMUB2020254), China.

Author information

Authors and Affiliations

  1. College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu Province, China

    Hui Huang, Jie Xue & Mei-Lin Xie

  2. Department of Neurosurgery, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhou, Jiangsu Province, China

    Tao Xie

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Contributions

HH and JX performed the animal and cell experiments. HH collected and analyzed the results and wrote the draft of the manuscript. JX revised and edited the manuscript. MLX and TX designed the experiments and reviewed the manuscript. All authors read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Mei-Lin Xie or Tao Xie.

Ethics declarations

Conflict of interest

H. Huang, J. Xue, M.-L. Xie and T. Xie declare that they have no competing interests.

Ethical standards

The animal study was conducted in accordance with the guidelines for the use and care of experimental animals of Soochow University and approved by the University Animal Ethics Committee (certificate no. 202103A0994; approval date 8 April 2021).

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Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Hui Huang and Jie Xue contributed equally to this study.

Data availability

Data will be made available from the corresponding authors upon reasonable request.

Supplementary Information

Fig. S1

The results of full-length sequencing of GSK-3β

Fig. S2

GSK-3β protein expression in pcDNA3.1(+)-GSK-3β plasmid-transfected HCC-LM3 cells (A) or SK-Hep-1 cells (B). Data are presented as the mean ± SD of three different independent experiments. **P < 0.01 vs. the pcDNA3.1(+)-NC group. There was no obvious difference between the control group and the pcDNA3.1(+)-NC group

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Huang, H., Xue, J., Xie, ML. et al. Osthole inhibits GSK-3β/AMPK/mTOR pathway-controlled glycolysis and increases radiosensitivity of subcutaneous transplanted hepatocellular carcinoma in nude mice. Strahlenther Onkol 200, 444–452 (2024). https://doi.org/10.1007/s00066-023-02173-8

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