Silencing LncRNA PVT1 Reverses High Glucose-Induced Regulation of the High Expression of PVT1 in HRMECs by Targeting miR-128-3p

Abstract

This paper aims to discuss the possibility of lncRNA PVT1 as a diagnostic biomarker for diabetic retinopathy (DR) and explore the underlying mechanism. Real-time quantitative polymerase chain reaction (RT-qPCR) was selected to determine the expression level of lncRNA PVT1 in the serum of all subjects. The receiver operating characteristic (ROC) curve reflected the diagnostic significance of PVT1 for DR patients. The Cell Counting Kit-8 (CCK-8) and Transwell assays were used to evaluate the effect of PVT1 expression on the proliferation and migration of human retinal microvascular endothelial cells (HRMECs). The luciferase reporter gene was selected to verify the interaction between PVT1 and miR-128-3p. The relative expression level of PVT1 in serum was higher in both the DB and DR group than in the healthy controls group (HC), and it was highest in the DR group. ROC curve indicated that serum PVT1 could distinguish between HC and DB patients, DB patients and DR patients, respectively. In vitro, high glucose induction significantly increased the proliferation and migration capabilities of HRMECs, but silencing PVT1 (si-PVT1) downregulated the proliferation and migration capabilities of HRMECs. The detection of luciferase reporter gene showed that lncRNA PVT1 targeted miR-128-3p, and there was a negative correlation in the serum of DR patients. In conclusion, this study confirmed that lncRNA PVT1 might regulate the process of DR by targeting miR-128-3p, and has the potential as a biomarker for the diagnosis of DR.

Downregulation of lncRNA ASMTL-AS1 in Epithelial Ovarian Cancer Correlates with Worse Prognosis and Cancer Progression

“…The miR-1228-3p mimic or inhibitor and the vectors (WT-ASMTL-AS1 or MUT-ASMTL-AS1) were co-transfected into SK-OV-3 and UACC-1598 cells. Luciferase activity was detected using Dual-Luciferase Reporter Assay System kit (Promega, USA) [20].…”

Diabetic Retinopathy: Are lncRNAs New Molecular Players and Targets?

“…Particularly, in diabetic patients with DR or in human retinal endothelial cells cultured with HG concentrations, the upregulation of several different lncRNAs has been recently reported. Although further analyses are needed, for most of the mentioned lncRNAs, preliminary evidence suggests a putative role in DR. Of note, the in vitro knockdown of these lncRNAs ameliorates retinal dysfunctions improving various related-biological processes, such as proliferation, apoptosis, migration, angiogenesis, inflammation, or redox state [ 62 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 , 126 ]. For instance, ARPE-19 cells exposed to HG display increased expression of IGF2 Antisense RNA ( IGF2-AS ) and enhanced apoptosis, and IGF2-AS silencing restrains apoptosis, inducing IGF2/Akt signaling and reducing Casp-9 expression [ 114 ].…”

“…Additionally, a recent work reports that the knockdown of TUG1 ameliorates diabetic retinal vascular dysfunction through regulating miR-524-5p/ FGFR2 axis [ 124 ]. Furthermore, independent studies in patients with DR showed the negative correlations between Plasmacytoma Variant translocation 1 ( PVT1 ) and miR-128-3p, as well as between the lncRNA OGRU and miR-320, supporting that also these lncRNAs may affect miRNA-mediated networks [ 62 , 125 ]. Particularly, in Müller cells cultured in HG conditions, OGRU suppression significantly restores miR-320 expression, and it represses the ubiquitin-specific peptidase 14 ( USP14 ) expression whereas, on the opposite, the upregulation of miR-320 reduces TGF-β1 signaling and impairs inflammation and oxidative stress [ 126 ].…”

Knockdown of lncRNA PVT1 protects human trabecular meshwork cells against H2O2-induced injury via the regulation of the miR-29a-3p/VEGF/MMP-2 axis

“…Additionally, starBase predicted the presence of a putative complementary sequence for miR-29a-3p on lncRNA plasmacytoma variant translocation 1 (PVT1). LncRNA PVT1 is highly expressed and acts as an oncogene in multiple cancers [ 23 ], and also functions in ophthalmic diseases such as diabetic retinopathy [ 24 ], retinoblastoma [ 25 ], and diabetic cataract [ 26 ]. However, the mechanism underlying the role of lncRNA PVT1 in glaucoma remains unknown.…”