Silencing of LncRNA PVT1 inhibits the proliferation, migration and fibrosis of high glucose-induced mouse mesangial cells via targeting microRNA-93-5p

Abstract

Objective: The present study aimed to investigate the regulatory role of long non-coding RNA plasmacytoma variant translocation 1 (PVT1) on 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. EdU and Colony formation, Annexin V-PI staining, Muse cell cycle, Scratch, and Transwell assays 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 enzyme-linked immunosorbent assay (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. The targeting relation between miR-93-5p and PVT1 was predicted by StarBase3.0 (an online software for analyzing the targeting relationship) and identified by Dual-luciferase reporter (DLR) 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.

Long Non-coding RNA: An Emerging Contributor and Potential Therapeutic Target in Renal Fibrosis

“…PVT1 knockdown inhibits cellular migration, invasion, proliferation, and fibrosis. Furthermore, PVT1 knockdown blocks the PI3K/Akt/mTOR signal and promotes the apoptosis of MMCs under HG conditions by upregulating miR-93-5p (Li J. et al, 2020). Furthermore, Zhong et al (2020) validated that silencing PVT1 might relieve HGinduced FN and α-SMA expression and proliferation in hMCs by inhibiting the NF-κB pathway via the miR-23b-3p/WT1 axis.…”

“…Furthermore, lncRNA/miRNA interactions are a common regulatory strategy in RF, but the gene regulatory network is complex and is yet to be fully elucidated. For example, PVT1 and XIST could bind to miR-93-5p at the predicted site; silencing of PVT1 or XIST inhibits the fibrosis process in HG-treated MMCs or HK-2 Li J. et al, 2020). Similarly, MALAT1 and MIAT could influence cell proliferation, viability, migration, and the EMT process by binding to miR-145 in TGF-β1-stimulated HK-2 cells (Liu B. et al, 2019;Liu et al, 2020;Wang Z. et al, 2020).…”

The DR1‑CSE/H2S system inhibits renal fibrosis by downregulating the ERK1/2 signaling pathway in diabetic mice

“…The effect of SKF38393 on cell viability, proliferation, proliferation-associated protein expression, fibrosis-associated protein expression and p-ERK1/2 activity was similar to that of PD98059 (Figs. [1][2][3][4][5][6]. This finding suggested that the DR1-CSE/H 2 S pathway inhibited MC dysfunction by inhibiting ERK1/2 activation.…”

“…In the presence of hyperglycemia and other stimulating factors, MCs proliferate abnormally and secrete a large amount of extracellular matrix, which is deposited in the mesangial area ( 5 ). MCs can migrate to the gap between glomerular capillary endothelial cells and the basement membrane and even protrude into the surrounding capillary cavity, resulting in capillary occlusion and ultimately leading to glomerulosclerosis ( 6 ). MC proliferation serves a key role in the process of DN, but the mechanism of MC proliferation in diabetic patients is complex.…”

Long noncoding RNA MIAT inhibits the progression of diabetic nephropathy and the activation of NF-κB pathway in high glucose-treated renal tubular epithelial cells by the miR-182-5p/GPRC5A axis

“…lncRNAs have also been regarded as important regulators in the regulation of DN [ 11 , 12 ]. PVT1 silence reduced cell proliferation, migration, and fibrosis in DN cell model [ 13 ]; NEAT1 acted as a pro-inflammatory molecule in DN [ 14 ]. Myocardial infarction-associated transcript (MIAT) downregulation has been correlated to the renal tubular epithelial injury in DN [ 12 ], and MIAT could enhance cell viability in high glucose (HG)-treated renal tubular cells by increasing the expression of Nrf2 [ 15 ].…”