Long Non-coding RNA MALAT1 Alleviates the Elevated Intraocular Pressure (Eiop)-induced Glaucoma Progression via Sponging miR-149-5p

ABSTRACT

Background: Glaucoma is an optic neuropathic disease and contributed to the irreversible blindness caused by the slow death of retinal ganglion cells (RGCs). Long non-coding RNA (lncRNA) metastasis associated lung adenocarcinoma transcript 1 (MALAT1) was reported to be aberrantly expressed in diverse diseases, including glaucoma. However, the mechanism of MALAT1 in glaucoma was still undefined.

Methods: The levels of MALAT1, microRNA-149-5p (miR-149-5p) in RGCs cultured under elevated pressure were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The putative target of MALAT1 was predicted by starBase v2.0 online database, and dual luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay were performed to verify this interaction. The cell viability of RGCs was measured by Cell Counting Kit-8 (CCK-8) assay. The apoptotic rate was evaluated via flow cytometry. The protein levels of apoptosis-related proteins (Bax, B-cell lymphoma 2 (Bcl-2)) and Cleaved caspase 3 were assessed by Western blot.

Results: The level of MALAT1 was significantly down-regulated, and the level of miR-149-5p was distinctly up-regulated in RGCs under pressure in a dose-dependent manner. Functionally, MALAT1 overexpression or miR-149-5p inhibitor alleviated the inhibitory effect on cell viability and the promoted effect on apoptotic rate of RGCs in EIOP. The interaction between MALAT1 and miR-149-5p was predicted by starBase v2.0 online database, and dual luciferase reporter assay, RIP assay and RNA pull-down assay validated the interaction. Combined with the loss and gain experiment results, miR-149-5p was negatively interacted with MALAT1. Furthermore, miR-149-5p mimics mitigated the promoted impact on cell viability and the suppressive impact on apoptotic rate by targeting MALAT1.

Conclusion: MALAT1 promoted cell proliferation and inhibited cell apoptosis of RGCs via targeting miR-149-5p in glaucoma in vitro, which might shed light on the mechanism of glaucoma pathogenesis.

KEYWORDS:

• lncRNA MALAT1
• miR-149-5p
• RGCs
• glaucoma

Acknowledgments

Not applicable.

Authors’ contribution

Conceptualization and Methodology: Jin Gong and Junling Wang; Formal analysis and Data curation: Jin Gong and, Jing Dan; Validation and Investigation: Linling Wang and Ping Wang; Writing-original draft preparation and Writing-review and editing: Linling Wang, Jin Gong and Jing Dan; Approval of final manuscript: all author.

Ethics approval and consent to participate

The present study was approved by the ethical review committee of the Affiliated Renhe Hospital of China Three Gorges University. Written informed consent was obtained from all enrolled patients.

Competing interests

The authors declare that they have no competing interests.

Availability of data and materials

The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.

Additional information

Funding

This study was hosted by the The Affiliated Renhe Hospital of China Three Gorges University (The Second Clinical Medical College of China Three Gorges University) and funded by Joint Fund of Hubei Provincial Health Commission (WJ2019H562).