miRNA expression profiling uncovers a role of miR‐302b‐3p in regulating skin fibroblasts senescence

Abstract

Numbers of emerging evidence suggest that variable microRNA (miRNA) expression facilitates the aging process. In this study, we distinguished aberrant miRNA expression in aged skin and explored the biological functions and potential mechanism of upregulated miR‐302b‐3p. At first, miRNA microarray analysis was examined to explore miRNA expression profiling in the skin of aging mice model by D‐galactose (d‐gal) injection. We identified 29 aberrant miRNAs in aged mice skin. Next, KEGG enrichment analysis was conducted with DIANA‐miPath v3.0, which was revealed that enrichment pathways involved in such processes as extracellular matrix‐receptor interaction, MAPK signaling pathway, and mammalian target of rapamycin (mTOR) signaling pathway. The target genes of deregulated miRNAs were predicted from four bioinformatic algorithms (miRDB, Targetscan, miRwalk, and Tarbase). The interaction network of miRNAs and their targets were visualized using Cytoscape software. As a result, we found that some hub genes (including JNK2, AKT1/2/3, PAK7, TRPS1, BCL2L11, and IKZF2) were targeted by 12 potential miRNAs (including miR‐302b‐3p, miR‐291a‐5p, miR‐139‐3p, miR‐467c‐3p, miR‐186‐3p, etc.). Subsequently, we identified five upregulated miRNA via quantitative polymerase chain reaction and all of them were confirmed increased significantly in aged skin tissues compared with young control tissues. Among them, high expression of miR‐302b‐3p was verified in both aged skin tissues and senescence fibroblasts. Furthermore, miR‐302b‐3p mimic accelerated skin fibroblast senescence and suppressed the longevity‐associated gene Sirtuin 1(Sirt1) expression, whereas miR‐302b‐3p inhibitor could delay skin fibroblast senescence and contribute Sirt1 expression. In addition, we demonstrated that c‐Jun N‐terminal kinase 2(JNK2) is a direct target of miR‐302b‐3p by a luciferase reporter assay. An inverse correlation was verified in fibroblasts between miR‐302b‐3p and JNK2. Most importantly, siRNA JNK2 confirmed that low expression of JNK2 could accelerate fibroblasts senescence. In conclusion, our results indicated that overexpressed miR‐302b‐3p plays an important biological role in accelerating skin aging process via directly targeting JNK2 gene.

MicroRNA-302b mitigates renal fibrosis via inhibiting TGF-β/Smad pathway activation

“…By binding to 3 0 -untranslated regions of mRNA, miRNAs can suppress translation of target genes (15). miR-302b has been proven to be involved in many important pathologic processes including suppressing lung cancer cell proliferation ( 16), inhibiting osteosarcoma cell invasion (17), and accelerating the skin aging process (18). Recently, more and more studies have focused on the interaction between miR-302 family members and renal cell fibrosis.…”

Growth factors‐based platelet lysate rejuvenates skin against ageing through NF‐κB signalling pathway: In vitro and in vivo mechanistic and clinical studies

“…To determine the anti‐ageing efficacy and explore the underlying mechanism of human PL (hPL) on skin, we retrospectively analysed subjects treated with hPL and then established an aged skin model of nude mice and HDFs by using d ‐galactose (D‐gal) for efficacy and mechanism study. D‐gal‐induced ageing model has been widely used to mimic intrinsic or natural ageing of human beings for anti‐ageing studies, which causes oxidative stress and cell senescence by triggering inflammatory processes on skin 36–39 . In this study, the efficacy and mechanism of hPL on skin ageing were determined for the first time, providing a promising therapeutic strategy for anti‐ageing treatment.…”

“…D‐gal‐induced ageing model has been widely used to mimic intrinsic or natural ageing of human beings for anti‐ageing studies, which causes oxidative stress and cell senescence by triggering inflammatory processes on skin. 36 , 37 , 38 , 39 In this study, the efficacy and mechanism of hPL on skin ageing were determined for the first time, providing a promising therapeutic strategy for anti‐ageing treatment.…”

Integrated microarray analysis of key genes and a miRNA‑mRNA regulatory network of early‑onset preeclampsia

“…For example, Huang et al (12) studied traumatic brain injury-related genes with gene expression microarray analysis. Tan et al (13) utilized a mirna expression microarray assay to identify miR-302b-3p as regulator of skin fibroblast senescence.…”