LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

Abstract

Long noncoding RNAs (lncRNAs) have been discovered to play a key role in adipogenesis, while the role of lncRNA human leukocyte antigen complex group 11 (HCG11) in adipocyte differentiation has not been studied clearly. We used human adipose-derived mesenchymal stem cells (hAdMSCs) to establish a model of cell differentiation in vitro and found that expression of lncRNA HCG11 was decreased during adipogenesis through real-time quantitative polymerase chain reaction analysis. Then, hAdMSCs were transfected with pcDNA-HCG11 or HCG11-shRNA (sh-HCG11); the adipogenic marker proteins were detected by Western blot, and the activity of lipogenesis enzymes was detected by spectrophotometry. The expression of CCAAT-enhancer-binding protein α, fatty acid-binding protein, peroxisome proliferator-activated receptor gamma 2 and the levels of acetyl coenzyme A carboxylase and fatty acid synthase FAS were significantly downregulated in hAdMSCs at different stages transfected with pcDNA-HCG11, while knockdown of lncRNA HCG11 promoted adipocyte differentiation. Bioinformatic analysis indicated that miR-204-5p was a potential target gene of HCG11, which was confirmed by luciferase reporter gene analysis and RNA pull-down analysis. In addition, miR-204-5p directly targeting the 3′-untranslated region of SIRT1 was also predicted by StarBase and verified by luciferase reporter gene analysis. Enforced expression of miR-204-5p negatively regulated the SIRT1 protein level. Furthermore, SIRT1 overexpression significantly inhibited adipogenic marker protein, levels of lipogenesis enzymes, and the proliferation of hAdMSCs. When pcDNA-HCG11 and miR-204-5p mimic were co-transfected into hAdMSCs, we found that the miR-204-5p mimic reversed the suppressor effect of pcDNA-HCG11. Taken together, we found that HCG11 negatively regulated cell proliferation and adipogenesis by the miR-204-5p/SIRT1 axis. Our findings might provide a new target for the study of adipogenesis in hAdMSCs and obesity.

Exosome derived from human adipose‐derived stem cell improve wound healing quality: A systematic review and meta‐analysis of preclinical animal studies

“…LncRNA XIST shuttled ADSC‐EV suppresses myocardial pyroptosis in atrial fibrillation through miR‐214‐3p‐mediated Arl2 51 . lncRNA HCG11 negatively regulates cell proliferation and adipogenesis by the miR‐204‐5p/SIRT1 23 . ADSC‐EV itself carries non‐coding RNAs that improve the quality of wound healing, and non‐coding RNAs can also be engineered and packaged in exosomes to act as messengers in intercellular according to the needs of patients.…”

“…The prospect of using EV enriched in such microRNA to improve the scarring process is now formed and opened. 20 , 21 In previous studies, the expression profiles of microRNA in cells reflect not only donor specificity but also that exosomes containing other non‐coding RNA (e.g., lncRNA, circleRNA) are influenced by differentiated cell types 22 , 23 , 24 ;Therefore, different biological properties can be expressed using exosomes delivering different cargo components.…”

Silencing Long Non-coding RNA Kcnq1ot1 Limits Acute Kidney Injury by Promoting miR-204-5p and Blocking the Activation of NLRP3 Inflammasome

“…Besides, cell proliferation was promoted, cell apoptosis was inhibited, and expressions of NLRP3, caspase1, and ASC were decreased in HK-2 cells transfected with oe-Kcnq1ot1 + miR-204-5p mimic compared with HK-2 cells transfected with oe-Kcnq1ot1 + mimic NC. miR-204-5p promotes cell proliferation in human adipose-derived mesenchymal stem cells ( Li D. et al, 2020 ). miR-204-5p could inhibit the inflammatory response in lipopolysaccharide-treated rat mesangial cells ( Chen et al, 2018 ).…”

Long Noncoding RNAs: Novel Important Players in Adipocyte Lipid Metabolism and Derivative Diseases

“…LncHCG11 is another target for inhibiting adipogenesis. Li et al (2020) found that the expression of lncHCG11 declines during the adipogenic differentiation in an in vitro hADMSCs differentiation model. Specifically, both the activity of related lipogenesis enzymes and the expressions of adipogenic proteins increase in HCG11 knockdowns, while the reverse response is observed when HCG11 is overexpressed.…”