LncRNA PLCD3-OT1 Functions as a CeRNA to Prevent Age-Related Cataract by Sponging miR-224-5p and Regulating PLCD3 Expression

Abstract

PURPOSE. Long noncoding RNAs (lncRNAs) are important in disease progression and cellular functions. This study aimed to conduct global lncRNA profiling and characterize the role of lncRNA 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase delta 3-sence RNA 1 (PLCD3-OT1) in the progression of age-related cataract (ARC). METHODS. We performed lncRNA expression profiling of lens capsule from ARC groups and age-matched groups using high-throughput RNA-sequencing. Real-time PCR was conducted to detect the expression pattern of lncRNA and mRNA in the clinical samples and cell model. Assays of cell-counting kit-8, 5 0-ethynyl-2 0-deoxyuridine, TUNEL, and propidium iodide staining were used to detect cell viability, proliferation, apoptosis, and cell cycle. We also performed fluorescence in situ hybridization assay to detect the location of lncRNA, and verified the endogenous competitive RNA mechanism between miRNAs, lncRNAs, and target genes via double-luciferase reporter analyses. RESULTS. The expression of lncRNA PLCD3-OT1 and PLCD3 were significantly decreased in ARC. PLCD3-OT1 overexpression promoted the expression of PLCD3, cell viability, proliferation, and inhibited cell apoptosis upon oxidative stress, while knockdown of PLCD3 showed the opposite results. Mechanistically, PLCD3-OT1functions through positively regulation the expression of PLCD3. In addition, PLCD3-OT1 may act as a ceRNA to regulate the expression of PLCD3 through competition for miR-224-5p. CONCLUSIONS. PLCD3-OT1 and PLCD3 may become potential therapeutic targets for the prognosis, diagnosis, and treatment of ARC.

Identification and Characterization of N6-Methyladenosine CircRNAs and Methyltransferases in the Lens Epithelium Cells From Age-Related Cataract

“… 2 Several studies have shown that there are strong links of the cellular and molecular pathogenesis of ARC to DNA damage, DNA repair, oxidative stress, proteolysis, ubiquitination, apoptosis and autophagy. 3 – 7 In particular, the repairability of DNA oxidation damage plays a crucial role in maintaining the lens’ transparency. 7 …”

“… 3 – 7 In particular, the repairability of DNA oxidation damage plays a crucial role in maintaining the lens’ transparency. 7 …”

“…Our previous studies reported the shreds of evidence of non-coding RNAs in the regulation of gene expression in lens epithelium cells (LECs). 7 − 10 Circular RNAs (circRNAs) are a novel class of noncoding RNAs, which are generated by the spliceosome via back splicing: the 3′ end of an exon is covalently linked to the 5′ end of an upstream exon. 11 , 12 Most circRNAs originate from protein-coding genes and contain complete exons.…”

The Emerging Roles of Non-Coding RNAs in Cataract

“…Emerging studies reveal that the dysregulation of lncRNAs are impact the development and progression of cataract [31] [32] [33] [34] [35]. Compared with transparent lenses, KCNQ1OT1, TUG1, and H19 has identified to have signifi-cantly increased expression in age-related cataract [36] [37] [38]. The significant differential expression of lncRNAs in age-related cataract human lenses indicates that lncRNAs might play a crucial role in cataract formation ( Table 2).…”

“…Therefore, H19 could be a useful marker in ARC. LncPLCD3-OT1 may act as a ceRNA to regulate the expression of PLCD3 by combining with mi-224-5p [38] and also plays a protective role in ARC [38]. Highly expressed TUG1 promotes lens epithelial cell apoptosis via sponging miR-421 [37].…”