LncRNA NEAT1 Promotes High Glucose-Induced Mesangial Cell Hypertrophy by Targeting miR-222-3p/CDKN1B Axis

Abstract

Glomerular hypertrophy is an early morphological alteration in diabetic nephropathy. Cyclin-Dependent Kinases have been shown to be required for high glucose (HG)-induced hypertrophy; however, the upstream regulators of CDKN1B in glomerular hypertrophy remain unclear. Herein we describe a novel pathway in which Long noncoding RNA (lncRNA) NEAT1 regulates the progression of mesangial cell hypertrophy via a competing endogenous RNA (ceRNA) mechanism. Real-time PCR was performed to detect the relative NEAT1 and miR-222-3p expressions and further confirmed the relationship between NEAT1 and miR-222-3p. Cell cycle was evaluated by flow cytometry. The related mechanisms were explored by Western blot, RNA immunoprecipitation and chromatin immunoprecipitation assay. We show that NEAT1 forms double stranded RNA (dsRNA) with miR-222-3p, thus limiting miR-222-3p’s binding with CDKN1B. This release of CDKN1B mRNA leads to elevated CDKN1B protein expression, resulting in hypertrophy. In addition, we demonstrated that STAT3 which is activated by HG induces the transcription of NEAT1 by binding to its promoter. Our findings underscore an unexpected role of lncRNAs on gene regulation and introduce a new mode of proliferation regulation in mesangial cells.

Long non-coding RNA L13Rik promotes high glucose-induced mesangial cell hypertrophy and matrix protein expression by regulating miR-2861/CDKN1B axis

“…G Wolf et al reported that HG promoted p27Kip1 expression in cultured mouse mesangial cells and may concubine to the process of diabetic glomerular hypertrophy [23]. Liao et al demonstrated that NEAT1 accelerated HG-induced mesangial cell hypertrophy by regulating miR-222-3p/CDKN1B axis [24]. In the current study, we found that CDKN1B was the direct target of miR-2861, as evidenced by the bioinformatics prediction, dual-luciferase-reporter gene and western blotting assay.…”

The Interplay of NEAT1 and miR-339-5p Influences on Mesangial Gene Expression and Function in Various Diabetic-Associated Injury Models

“…With regard to DKD, others showed a positive and negative correlation, respectively, between elevated urinary NEAT1 and various podocyte damage markers like synaptopodin and podocalyxin or GFR [ 42 ]. HG-exposed proximal tubular cells (HK-2) or murine MCs (mMCs) exhibited an increase in NEAT1 expression in a dose- or time- dependent manner [ 31 , 43 ]. Contrarily to others, our data indicate that NEAT1 expression is not affected by HG using an osmotic control and a species model, in which NEAT1 is annotated (human or murine) [ 31 , 43 , 44 ].…”

Long non-coding RNA L13Rik promotes high glucose-induced mesangial cell injury by regulating miR-2861/CDKN1B axis

“…In fact, more and more studies have revealed that lncRNAs function as its biological role in this way. In the study we demonstrated that L13Rik acts as a ceRNA to sponge miR-2861, resulting in the de-repression of its target CDKN1B, a gene known to accelerate MC injury (Liao et al, 2020).…”

“…The cell-cycle arrest is a hallmark of MC hypertrophy (Shankland, 1999), and cyclin-dependent kinase inhibitor 1A (CDKN1A), a kind of cyclin-dependent kinase inhibitor, is required for glomerular hypertrophy in experimental DN (Al-Douahji et al, 1999). In addition, up-regulated CDKN1B by lncRNA-NEAT1 promotes MC hypertrophy (Liao et al, 2020). In the present study, we demonstrated that L13Rik was significantly increased in DN patients and rats, and L13Rik depletion effectively reduced HG-induced MC survival, ECM accumulation, and cell hypertrophy through sponging miR-2861 and thus de-repressing its target CDKN1B.…”