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Exosomal microRNA-222-3p increases UVB sensitivity of lens epithelium cells by suppressing MGMT

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Abstract

Background

Age-related cataract (ARC) is a leading cause of blindness worldwide with multiple pathogenic factors. Oxidative damage of lens epithelium cells (LECs) is one of the well-accepted pathogenesis of ARC which can be regulated by DNA repair genes (DRGs). The present research aimed to clarify the regulatory mechanism of exosomal microRNAs (miRNAs) on DRGs in LECs.

Methods

The LECs oxidative damage model was established by UVB-irradiation on SRA01/04 (human lens epithelium cell line). Exosomes from UVB-irradiated cells (UVB-exo) and exosomes from normal control cells (NC-exo) were collected from the culture medium. To explore the functions of LECs exosomes, SRA01/04 were incubated with UVB-exo/NC-exo. Then, we detected SRA01/04 proliferation, viability and apoptosis respectively using 5′-ethynyl-2′-deoxyuridine (EdU), cell-counting kit-8 (CCK-8) and TdT-mediated dUTP Nick-End Labeling (TUNEL) assay. Next, the miRNA expression profiles of UVB-exo and NC-exo were identified by miRNA microarrays. RNA expression in exosomes, cells, and clinical samples was verified by qRT-PCR. The location and expression of MGMT and CD63 proteins were detected by immunofluorescence and western blot. The 3′UTR regulation of miR-222-3p to MGMT was verified by luciferase analyses.

Results

MGMT down-regulated while miR-222-3p up-regulated in LECs sub-central anterior capsule from ARC lenses. MGMT and miR-222-3p expressions in central and peripheral LECs from anterior lens capsules were differential. UVB-exo can transport the up-regulated miR-222-3p from oxidative-damaged LECs to normal LECs, which could suppress MGMT expression and increase UVB sensitivity of LECs.

Conclusions

Findings on exosomal miRNA functions provided novel insights into pathogenesis of ARC. Exosomal miR-222-3p can be a potential target for prevention and cure of ARC.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This study was funded by the National Natural Science Foundation of China (Grant Nos.: 81974129, 82171038, 82101101), and the Science and Technology Project of Nantong Municipality (Grant No.: JC2019078).

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Author notes

  1. Jiawei Luo, Pengfei Li and Lihua Kang contributed equally to this work.

Authors and Affiliations

  1. Eye Institute, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong, 226001, Jiangsu, China

    Jiawei Luo, Pengfei Li, Lihua Kang, Min Ji, Tianqiu Zhou, Bai Qin, Junfang Zhang & Huaijin Guan

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Contributions

JL, LK and PL contributed equally to this work. All authors participated in experiments performing, data analysis and manuscript review before submission.

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Correspondence to Huaijin Guan.

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Luo, J., Li, P., Kang, L. et al. Exosomal microRNA-222-3p increases UVB sensitivity of lens epithelium cells by suppressing MGMT. Int Ophthalmol 43, 1611–1628 (2023). https://doi.org/10.1007/s10792-022-02560-7

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