釉基质蛋白诱导MC3T3-E1细胞成骨分化过程中微小核糖核酸的表达

发布时间：2018-07-05 06:59

本文选题：微小核糖核酸 + 成骨分化　；参考：《口腔材料器械杂志》2014年04期

【摘要】：目的探讨微小核糖核酸(micro ribonucleicacid,mi RNA)是否参与到釉基质蛋白(Enamel Matrix Derivative,EMD)诱导的小鼠前成骨细胞系MC3T3-E1细胞成骨分化的过程,并对发生显著变化的mi RNA进行分析。方法将MC3T3-E1用含EMD(刺激组)/不含EMD(对照组)的培养液培养0天、7天和14天,检测碱性磷酸酶(alkaline phosphatase,ALP)活性,实时聚合酶链式反应(RT-PCR)技术分析成骨分化标记物ALP、骨涎蛋白(bone sialoprotein,BSP)和骨钙素(osteocalcin,OC)的信使RNA(m RNA)水平的表达变化。利用基因芯片技术分析mi RNA的相对表达。结果 0天、7天、14天的ALP染色结果显示随着培养时间的增加,两组ALP活性均逐渐增强,EMD刺激组ALP活性增强较对照组更为明显。RT-PCR结果表明,与对照组相比,ALP、BSP、OC的m RNA表达量均显著增加,ALP与OC均于14天时表达量达到峰值,BSP则于7天时处于峰值表达,EMD刺激组MC3T3-E1成骨活性更强;各期11个mi RNA表达上调,28个mi RNA表达下调,其中mi R-335-5p,mi R-503已被证实可参与促进骨形成,而mi R-30家族(mi R-30a,-30b,-30c和-30d)则被证实参与抑制成骨。结论 mi RNA参与EMD诱导的MC3T3-E1细胞的成骨分化过程,这一发现可以为了解EMD促进成骨分化的机理及临床应用提供指导。
[Abstract]:Objective to investigate the role of micro ribonucleic acid (micro) in the process of osteogenic differentiation of mouse preosteogenic cell line MC3T3-E1 induced by Enamel Matrix Derivatives. Methods MC3T3-E1 was incubated with EMD / EMD (control group) for 7 and 14 days, and the activity of alkaline phosphatase was measured. The expression of ALP, bone sialoprotein and osteocalcin OC mRNA was analyzed by real-time polymerase chain reaction (RT-PCR). The relative expression of mi RNA was analyzed by gene chip technique. Results the results of ALP staining on day 0 and day 7 and day 14 showed that ALP activity increased gradually in both groups with the increase of culture time. The results of RT-PCR showed that ALP activity in EMD stimulated group was more obvious than that in control group. Compared with the control group, the mRNA expression of BSPOC was significantly increased. Both ALP and OC reached the peak at 14 days. The osteogenic activity of MC3T3-E1 stimulated by EMD at 7 days was higher than that of the control group, and the expression of 11 mi RNA was up-regulated and 28 mi RNA was down-regulated in each stage. Among them, mi R-335-5pnmmi R-503 has been proved to be involved in promoting bone formation, while the mi R-30 family (mi R-30aan-30ban-30c and -30d) has been proved to be involved in the inhibition of osteogenesis. Conclusion mi RNA is involved in the osteogenic differentiation of MC3T3-E1 cells induced by EMD, which may provide guidance for understanding the mechanism and clinical application of EMD in promoting osteogenic differentiation.
【作者单位】：上海交通大学医学院附属第九人民医院口腔修复科上海市口腔医学重点实验室;唾液腺疾病研究中心首都医科大学口腔医学院牙齿再生与功能重建重点实验室;
【基金】：国家自然科学基金资助项目(81170988) 上海市科学技术委员会基金资助项目(11ZR1420200)
【分类号】：R329.2

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