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Integrin β6/Annexin A2 axis triggers autophagy to orchestrate hepatocellular carcinoma radioresistance

Cell Death & Differentiation volume 32, pages 689–701 (2025)Cite this article

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Abstract

Radiotherapy (RT) is one of the main therapies for hepatocellular carcinoma (HCC), but its effectiveness has been constrained due to the resistance effect of radiation. Thus, the factors involved in radioresistance are evaluated and the underlying molecular mechanisms are also done. In this present study, we identified Integrin β6 (ITGB6) as a potential radioresistant gene through an integrative analysis of transcriptomic profiles, proteome datasets and survival using HCC cases treated with IR. We show that ITGB6 functionally contributed to radioresistance by activating autophagy through a series of in vitro and in vivo methods, such as clonogenic assays, autophagy flux (LC3B-GFP-mCherry reporter) analysis and a subcutaneous transplantation model. Mechanically, ITGB6 binds to Annexin A2 (ANXA2) and enhanced its stability by competitively antagonizing proteasome mediated ANXA2 degradation, thereby promoting autophagy and radioresistance. Notably, HCC radioresistance was significantly improved by either blocking ITGB6 or autophagy, but the combination was more effective. Importantly, ITGB6/ANXA2 axis triggered autophagic program endowed HCC cells with radioresistant activity in a radiated patient-derived xenograft (PDX) model and hydrodynamic injection in liver-specific Itgb6-knockout mice, further supported by clinical evidence. Together, our data revealed that ITGB6 is a radioresistant gene stabilizing the autophagy regulatory protein ANXA2, providing insights into the biological and potentially clinical significance of ITGB6/ANXA2 axis in radiotherapy planning of HCC.

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Fig. 1: Ionizing radiation (IR)-mediated ITGB6 overexpression is clinically relevant in HCC radioresistance.
Fig. 2: ITGB6 confers radioresistance to HCC cells probably through autophagy in vitro and in vivo.
Fig. 3: ITGB6 induces autophagic flux of HCC cells in vitro.
Fig. 4: ANXA2 is essentially required for ITGB6-induced autophagy and radioresistance in HCC.
Fig. 5: ITGB6 interacts with ANXA2 to abrogate ANXA2 ubiquitylation.
Fig. 6: ITGB6/ANXA2 axis guides the applicability of HCC patients to RT.
Fig. 7: Working model depicting the mechanisms that ITGB6/ANXA2 axis and autophagy underlying radioresistance of HCC cells.

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Data availability

RNA-seq data was deposited in the NCBI SRA database with the BioProject accession PRJNA1170233. Full blots of immunoblot experiments can be found in supplemental materials. All other data are available from the corresponding authors upon request.

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Acknowledgements

We acknowledge Dr. Min-shan Chen and Dr. Yao-jun Zhang, Department of Hepatobiliary Oncology, SYSUCC, for their help in collecting HCC samples. We thank the specimen donors at SYSUFAH and SYSUCC. The graphical abstract and working model were created with BioRender (http://biorender.com/).

Funding

The study was supported by the National Natural Science Foundation of Guangdong Province of China (No. 2022B1515020010) and the Young Scientists Fund of the National Natural Science Foundation of China (No. 82103771).

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Author notes

  1. These authors contributed equally: Ying Gao, Guangyan Wei, Hua Yu, Shuping Li.

Authors and Affiliations

  1. Department of Radiation Oncology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China

    Ying Gao, Guangyan Wei, Shuping Li, Yuhao Tang, Xin Yue & Yong Chen

  2. Cancer Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China

    Ying Gao, Guangyan Wei, Shuping Li, Yuhao Tang, Xin Yue & Yong Chen

  3. Laboratory of General Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China

    Ying Gao & Shuping Li

  4. School of Life Sciences, Guangzhou University, Guangzhou, Guangdong, China

    Hua Yu

  5. Department of Liver Surgery, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong, China

    Yuhao Tang

  6. Institute of Precision Medicine, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China

    Xin Yue

  7. Department of Interventional Medicine, Guangzhou First Pepople’s Hospital, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, China

    Meixiao Zhan

  8. Guangdong Provincial Key Laboratory of Tumor Interventional Diagnosis and Treatment, Zhuhai People’s Hospital (Zhuhai Clinical Medical College of Jinan University), Zhuhai, Guangdong, China

    Meixiao Zhan

  9. Center of Hepato-Pancreato-Biliary Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China

    Jian Wu

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Contributions

JW and MXZ provided the study concept and design. YG, GYW and HY interpreted and analyzed the data. YG, GYW, HY and SPL performed the experiments. YG, GYW, YHT, XY, and YC collected the patients’ samples. YG, GYW, MXZ and JW wrote the manuscript. JW and MXZ supervised the study and reviewed the manuscript. All authors approved the final version of the manuscript.

Corresponding authors

Correspondence to Meixiao Zhan or Jian Wu.

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Competing interests

The authors declare no competing interests.

Ethics approval

The study was conducted in accordance with the Declaration of Helsinki. The informed consent was obtained from all participants. For the use of clinical materials, the study was approved by the Ethics Committee of the First Affiliated Hospital of Sun Yat-sen University (SYSUFAH) (Approval No. [2022]438) and the Ethics Committee of Sun Yat-sen University Cancer Center (SYSUCC) (Approval No. B2019-008-01). Approval No. L102012019060I was granted by the Institutional Animal Care and Use Committee of Sun Yat-sen University Cancer Center for all mouse care and experiments. All methods were performed in accordance with the relevant guidelines and regulations in the present study.

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Gao, Y., Wei, G., Yu, H. et al. Integrin β6/Annexin A2 axis triggers autophagy to orchestrate hepatocellular carcinoma radioresistance. Cell Death Differ 32, 689–701 (2025). https://doi.org/10.1038/s41418-024-01411-5

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