Silencing of Long non-coding RNA Plasmacytoma variant translocation 1 inhibits the proliferation, migration, invasion and fibrosis of high glucose-induced mouse mesangial cells via targeting microRNA-93-5p

Abstract

Background: This study aimed to investigate the regulatory role of Long non-coding RNA plasmacytoma variant translocation 1 (PVT1) on the biological processes of high glucose (HG)-induced mouse mesangial cells (MMCs). Methods: PVT1 expression in diabetic nephropathy (DN) mice and HG-induced MMCs was detected by qRT-PCR. PVT1 was silenced in HG-induced MMCs by the transfection of PVT1 siRNA (si-PVT1). EdU and Colony formation, Annexin V-PI staining, Muse cell cycle, Scratch, and Transwell assay were performed to detect the cell proliferation, apoptosis, cell cycle, migration, and invasion, respectively. The contents of fibrosis factors in cell-culture supernatants were detected by ELISA. Western blot was performed to detect the expression of factors involved in apoptosis, cell cycle, migration and invasion, fibrosis, and PI3K/Akt/mTOR pathway. In addition, the targeting relation between miR-93-5p and PVT1 was predicted by StarBase3.0 and identified by Dual-luciferase reporter gene assay. Results: PVT1 was overexpressed in DN kidney tissues and HG-induced MMCs. HG-induced MMCs exhibited significantly increased EdU positive cells, cell colonies, S and G2/M phase cells, migration and invasion ability, and contents of fibrosis factors, as well as significantly decreased apoptosis rate compared with NG-induced MMCs. HG significantly up-regulated Bcl-2, CyclinD1, CDK4, N-cadherin, vimentin, Col. IV, FN, TGF-β1 and PAI-1, and down-regulated Bax, cleaved caspase-3, cleaved PARP, and E-cadherin in MMCs. Silencing of PVT1 eliminated the effects of HG in MMCs and blocked PI3K/Akt/mTOR pathway. MiR-93-5p was a target of PVT1, which eliminated the effects of PVT1 on HG-induced MMCs. Conclusions: PVT1 silencing inhibited the proliferation, migration, invasion and fibrosis, promoted the apoptosis, and blocked PI3K/Akt/mTOR pathway in HG-induced MMCs via up-regulating miR-93-5p.