The role and possible mechanism of lncRNA U90926 in modulating 3T3-L1 preadipocyte differentiation

Abstract

Background:Obesity is a risk factor for metabolic diseases, while preadipocyte differentiation or adipogenesis is closely related to obesity occurrence. Long noncoding RNAs (lncRNAs) are a unique class of transcripts in regulation of a variety of biological processes. Using cDNA microarray, we found lncRNA U90926 is negatively correlated with 3T3-L1 preadipocyte differentiation.Objective:The aim of this study was to explore the role of lncRNA U90926 (lnc-U90926) in adipogenesis and the underlying mechanisms.Methods:Quantitative real-time PCR (qPCR) was performed to determine lnc-U90926 expression in 3T3-L1 preadipocytes, differentiated adipocytes, and in adipose tissues form mice. RNA fluorescent in situ hybridization (FISH) was performed to determine the localization of lnc-U90926 in 3T3-L1 preadipocytes. The effects of lnc-U90926 on 3T3-L1 adipogenesis were analyzed with lentivirus-mediated gain- and loss-of-function experiments. Lipid accumulation was evaluated by oil red O staining; several adipogenesis makers were analyzed by qPCR and western blotting. Dual luciferase assay was applied to explore the transactivation of target genes modulated by lnc-U90926. All measurements were performed at least for three times.Results:Lnc-U90926 expression decreased along the differentiation of 3T3-L1 preadipocytes. In mice, lnc-U90926 is predominantly expressed in adipose tissue. Obese mice have lower lnc-U90926 expression in subcutaneous and visceral adipose tissue than non-obese mice. FISH results showed that lnc-U90926 was mainly located in the cytoplasm. Overexpression lnc-U90926 attenuated 3T3-L1 adipocyte differentiation as evidenced by its ability to inhibit lipid accumulation, to decrease the mRNA levels of peroxisome proliferator-activated receptor gamma 2 (PPARγ2), fatty acid binding protein 4 (FABP4) and adiponectin (AdipoQ) as well as to reduce the protein levels of PPARγ and FABP4 (P<0.05). Knockdown of lnc-U90926 showed opposite effects, which increased mRNA expression of PPARγ2, FABP4, CCAAT/enhancer-binding proteinα (C/EBPα) and AdipoQ.Conclusion:Lnc-U90926 attenuates 3T3-L1 adipocyte differentiation via inhibiting the transactivation of PPARγ2 or PPARγ.

The Construction and Analysis of the Aberrant lncRNA-miRNA-mRNA Network in Adipose Tissue from Type 2 Diabetes Individuals with Obesity

“…Despite their lack of protein coding capacity, there is increasing evidence that lncRNAs are widely involved in the regulation of gene expression. lncRNAs are generally considered to be important regulators of adipogenesis and adipocyte metabolism [13][14][15][16][17]. lncRNAs also play a role in the pathogenesis of T2DM and T2DM-related complications [18][19][20][21][22][23][24].…”

Long noncoding RNA Plnc1 controls adipocyte differentiation by regulating peroxisome proliferator‐activated receptor γ

“…Several lncRNAs have been found to regulate adipogenesis through modulating the expression of PPAR-g2. Lnc-U90926 suppressed 3T3-L1 adipocyte differentiation through inhibiting the transactivation of the PPAR-g2 promoter (43). lncRNA NEAT1 works with SRp40 to regulate PPAR-g2 splicing during adipogenesis in 3T3-L1 cells (44).…”

Cachexia‐related long noncoding RNA, CAAlnc1, suppresses adipogenesis by blocking the binding of HuR to adipogenic transcription factor mRNAs

“…ADNCR (adipocyte differentiation‐associated long noncoding RNA) represses adipogenesis by functioning as a competing endogenous RNA . Recently, AdipoQ AS lncRNA and lnc‐U90926 were also reported to play inhibitory roles in adipogenesis . Although numerous lncRNAs are known as regulators of adipogenesis, knowledge of lncRNAs that selectively modulate adipose metabolism during CAC development remains limited …”

“…22 Recently, AdipoQ AS lncRNA and lnc-U90926 were also reported to play inhibitory roles in adipogenesis. 23,24 Although numerous lncRNAs are known as regulators of adipogenesis, knowledge of lncRNAs that selectively modulate adipose metabolism during CAC development remains limited. 25 In our study, we aimed to identify novel and functionally important lncRNAs involved in CAL using mouse models.…”