miR-30a在雷帕霉素刺激人炎症牙周膜干细胞后的表达情况及其与Beclin1基因的靶向调控作用的研究

发布时间：2018-10-05 13:26

【摘要】：目的:研究雷帕霉素对人牙周膜干细胞中miR-30a的表达的影响,并验证其与Beclin1的靶向调控关系,探讨miR-30a与自噬途径在牙周膜干细胞的炎症作用机制。方法:分离培养人牙周膜干细胞,并分别进行50μg/L雷帕霉素作用2h,10nmol/L 3-甲基腺嘌呤(3-MA)作用12h,收集细胞,提取总RNA;应用实时荧光定量PCR(qRT-PCR)检测miR-30a的表达水平;应用qRT-PCR、Western blot及双荧光素酶报告系统验证miR-30a与Beclin1的相互作用关系。结果:雷帕霉素刺激细胞后,细胞内的miR-30a的表达升高;qRT-PCR、Western blot及双荧光素酶报告系统证实,miR-30a对Beclin1的mRNA及蛋白均有抑制,且与Beclin1的3’UTR具有靶向抑制作用。结论:miR-30a可靶向抑制自噬相关基因Beclin1的表达,并参与人牙周膜干细胞的细胞自噬过程。
[Abstract]:Aim: to investigate the effect of rapamycin on the expression of miR-30a in human periodontal ligament stem cells (PDSCs), and to verify the relationship between rapamycin and Beclin1, and to explore the inflammatory mechanism of miR-30a and autophagy pathway in periodontal ligament stem cells (PDSCs). Methods: human periodontal ligament stem cells were isolated and cultured, and treated with 50 渭 g / L rapamycin for 2 h or 10 nmol / L 3-methyladenine (3-MA) for 12 h. The total RNA; was collected and the expression of miR-30a was detected by real-time fluorescence quantitative PCR (qRT-PCR). QRT-PCR,Western blot and double luciferase report system were used to verify the interaction between miR-30a and Beclin1. Results: after stimulated by rapamycin, the expression of miR-30a in the cells was increased. The expression of QRT-PCR blot and double luciferase report system confirmed that pmiR-30a could inhibit the mRNA and protein of Beclin1, and had a targeted inhibitory effect with 3'UTR of Beclin1. Conclusion: 1 miR-30a can inhibit the expression of autophagy related gene Beclin1 and participate in the autophagy process of periodontal ligament stem cells.
【作者单位】：首都医科大学附属北京康复医院口腔科;北京大学口腔医院综合科;
【分类号】：R781.4

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