miR-21 promotes radiation-induced pulmonary fibrosis by targeting CREBL2

doi:10.3877/cma.j.issn.1674-6902.2025.02.003

Abstract

Abstract:

Objective

Dysfunction of type Ⅱalveolar epithelial cells （AT2） is considered one of the key pathological processes underlying the development of radiation-induced pulmonary fibrosis （RIPF）. miR-21 regulates epithelial to mesenchymal transition （EMT） during RIPF. However，the target gene through which miR-21 acts upon in the regulation of EMT remains to be confirmed. This study primarily investigates the molecular mechanisms and critical roles of miR-21 and CREBL2 in RIPF.

Methods

The chests of wild type（WT） mice and miR-21^-/- mice were irradiated with 15Gy cobalt source γ-rays to establish a RIPF model.Through living lung function examination and lung hydroxyproline detection to evaluate the degree of pulmonary fibrosis in irradiated mice. Establishment of the rat type Ⅱalveolar epithelial cells line （RLE-6TN） irradiation model，the RT-qPCR and WB were used to detect the expression of endoplasmic reticulum stress （ERS） and EMT related markers in irradiated cells. Differentially expressed genes were identified through a microarray screen and validated using a dual-luciferase reporter assay.

Results

After irradiation，WT mice exhibited a 14-fold increase in miR-21 expression at 8 weeks post-irradiation. Additionally，hydroxyproline expression increased by more than 2-fold，while CREBL2 expression decreased by 50%. In contrast，miR-21^-/- mice demonstrated an approximately 3-fold increase in CREBL2 expression following irradiation. The miR-21^-/- mice exhibited a reduced degree of lung fibrosis compared to WT mice. The markers of ERS and EMT in the lung tissue of miR-21^-/- mice were significantly reduced. Dual-luciferase reporter assays confirmed the specific binding of miR-21 to CREBL2.

Conclusion

Radiation-induced increase of miR-21 and decreased expression of its target gene CREBL2 in AT2 cells further activated the ERS signaling pathway，mediated EMT in AT2 cells，and finally led to the formation of RIPF.

Key words: Radiation induced pulmonary fibrosis, Epithelial to mesenchymal transition, miR-21, cAMP responsive element binding protein like 2, Endoplasmic reticulum stress

Jie Li, Yonghong Ran, Yuhui Hao. miR-21 promotes radiation-induced pulmonary fibrosis by targeting CREBL2[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(02): 220-225.

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URL: https://zhfbjbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-6902.2025.02.003

https://zhfbjbzz.cma-cmc.com.cn/EN/Y2025/V18/I02/220

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References 37

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