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Hypoxia promotes epithelial–mesenchymal transition in lung cancer cells via regulating the NRF2/miR‑27a/BUB1 pathway

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Abstract

Purpose

Lung cancer (LC) is the most common malignancy in the world. It is well that hypoxia is common in lung cancer, which contributes to lung cancer progression and metastasis [1]. miRNA-27a as a repressor factor is a lowly expression within non-small cell lung cancer (NSCLC). However, the molecular mechanism between miR-27a and hypoxia in lung cancer progression remains poorly understood. This study aims to explore hypoxia promotes epithelial-mesenchymal transition in lung cancer cells via regulating the NRF2/miR‑27a/BUB1 pathway.

Methods

We detect the expression of miR-27a after exposure to hypoxia conditions in lung cancer cells via qPCR. Using MTT assay and colony assay to assess the ability of proliferation in lung cancer cells under hypoxia or transfect miR-27a mimics. The capability of migration and invasion was evaluated by wound healing assay and Boyden-chamber assay. The mRNA and protein expression of EMT markers was respectively detected by qPCR and western blot. We detected NRF2 occupancy at the miR-27a promoter by ChIP-Seq analysis. Meanwhile, the luciferase assay verified BUB1 as a direct target of miR-27a.

Results

We found hypoxia promotes lung cancer cell proliferation, migration, invasion, and the epithelial-mesenchymal transition (EMT) process by inhibiting the miR-27a expression. miR-27a mimics significantly reduced the promotion effect of hypoxia on the invasion and proliferation of lung cancer cells. NRF2 as regulating the oxidation/anti-oxidation factor was activated under hypoxia conditions. The activation of NRF2 repressed miR-27a expression. On the contrary, the inhibitory effect of hypoxia on miR-27a was reversed when the NFE2L2 gene was silenced. Ectopic expression of NRF2 inhibited miR-27a expression under normoxia. We further validated BUB1 as a direct target of the miR-27a by luciferase assay.

Conclusion

Hypoxia promotes invasion and epithelial-mesenchymal transition of Lung cancer cells by regulating the NRF2/miR-27a/BUB1 axis.

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Funding

This work was funded by Sichuan Medical Association (Q18010, 2018-ZRQN-093).

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Authors and Affiliations

  1. Department of Respiratory and Critical Care Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan, People’s Republic of China

    Chunfeng Liu, Jun Deng & Songping Wang

  2. Inflammation and Allergy Diseases Research Unit, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan, People’s Republic of China

    Chunfeng Liu, Jun Deng & Songping Wang

  3. Faculty of Medicine, University of Munich, 80336, Munich, Germany

    Chunfeng Liu

  4. Department of General Surgery (Gastrointestinal Surgery), The Affiliated Hospital of Southwest Medical University, Taiping Str. 25, Luzhou, 646000, Sichuan, People’s Republic of China

    Lei Ren

  5. Department of Surgery, Klinikum Rechts der Isar, School of Medicine, Technical University of Munich, Ismaninger Str. 22, 81675, Munich, Germany

    Lei Ren

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  1. Chunfeng Liu
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Correspondence to Lei Ren.

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Authors declare that there is no conflict of interest in this study.

Ethical approval

This project has been approved by Southwest Medical University for experimental animal ethics. The approval number is:20211009–032.

Human and animal rights

The study was approved by the Southwest Medical University ethics committee. This article does not contain any studies with human participants or animals performed by any of the authors.

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Liu, C., Deng, J., Wang, S. et al. Hypoxia promotes epithelial–mesenchymal transition in lung cancer cells via regulating the NRF2/miR‑27a/BUB1 pathway. Clin Transl Oncol 25, 510–522 (2023). https://doi.org/10.1007/s12094-022-02965-x

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  • DOI: https://doi.org/10.1007/s12094-022-02965-x

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