miR-29c-3p promotes senescence of human mesenchymal stem cells by targeting CNOT6 through p53-p21 and p16-pRB pathways.
miR-29c-3p promotes senescence of human mesenchymal stem cells by targeting CNOT6 through p53-p21 and p16-pRB pathways.
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DOI：
10.1016/j.bbamcr.2016.01.005
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发表时间：
2016-04
期刊：
影响因子：
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通讯作者：
J. Shang;Yuan Yao;Xin Fan;L. Shangguan;J. Li;Huan Liu;Yue Zhou
中科院分区：
文献类型：
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作者：
J. Shang;Yuan Yao;Xin Fan;L. Shangguan;J. Li;Huan Liu;Yue Zhou
间充质干细胞是组织工程的重要种子细胞，也是细胞治疗的重要靶点。然而，MSCs在体外培养过程中的复制性衰老限制了其研究和临床应用。骨髓间充质干细胞复制性衰老的分子机制尚未完全阐明。有证据表明miRNAs在复制性衰老中起重要作用。微阵列分析发现，miR-29 c-3 p水平在MSC衰老过程中显著增加。在我们的研究中，我们研究了miR-29 c-3 p在MSC衰老中的作用。我们长期培养MSCs，上调和下调miR-29 c-3 p在MSCs中的表达，并检测衰老表型的变化。miR-29 c-3 p的过表达导致衰老相关β-半乳糖苷酶（SA-β-gal）染色增强、衰老相关分泌表型（SASP）、衰老相关异染色质灶（SAHF）、增殖能力降低、成骨分化延迟以及衰老标志物的相应变化，而miR-29 c-3 p的下调则导致相反的结果。双荧光素酶报告基因分析表明hocT 6是miR-29 c-3 p的靶基因。敲除hocT 6证实了其对MSC衰老的抑制作用。Western blot结果显示，miR-29 c-3 p诱导的MSCs衰老过程中，p53-p21和p16-pRB通路均被激活。总之，我们的研究结果表明，miR-29 c-3 p通过p53-p21和p16-pRB通路靶向hocT 6促进MSC衰老，并突出了转录后调控对干细胞衰老的贡献。
Mesenchymal stem cells (MSCs) are important seed cells for tissue engineering and are promising targets for cell-based therapies. However, the replicative senescence of MSCs duringin vitroculture limits their research and clinical applications. The molecular mechanisms underlying the replicative senescence of MSCs are not fully understood. Evidence suggests that miRNAs play important roles in replicative senescence. A microarray analysis found that the miR-29c-3p level was significantly increased during the MSC senescence process. In our study, we investigated the roles of miR-29c-3p in senescence of MSCs. We cultured MSCs for long periods of time, up and down-regulated the miR-29c-3p expression in MSCs, and examined the senescent phenotype changes. The over-expression of miR-29c-3p led to enhanced senescence-associated-β-galactosidase (SA-β-gal) staining, senescence associated secretory phenotype (SASP), senescence associated heterochromatic foci (SAHF), reduced proliferation ability, retarded osteogenic differentiation and corresponding changes in senescence markers, whereas the miR-29c-3p down-regulation had the opposite results. Dual-luciferase reporter assays demonstrated that CNOT6 is the target gene of miR-29c-3p. Knockdown of CNOT6 confirmed its inhibitory effects on the senescence of MSCs. In addition, Western blot results showed that both the p53–p21 and the p16–pRB pathways were activated during the miR-29c-3p-induced senescence of MSCs. In conclusion, our results demonstrate that miR-29c-3p promotes the senescence of MSCs by targeting CNOT6 through p53–p21 and p16–pRB pathways and highlight the contribution of post-transcriptional regulation to stem cell senescence.