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浙江大学学报（医学版）
浙江大学学报（医学版）  2016, Vol. 45 Issue (6): 575-580    DOI: 10.3785/j.issn.1008-9292.2016.11.03
骨组织代谢与再生专题     
微RNA-705对MC3T3-E1细胞成骨分化能力的影响
杨晓红1, 杨琨2, 廖立3, 金岩3
1. 遵义医学院附属口腔医院口腔修复科, 贵州 遵义 563000;
2. 遵义医学院附属口腔医院牙周科, 贵州 遵义 563000;
3. 第四军医大学口腔医院组织工程研究中心 军事口腔医学国家重点实验室, 陕西 西安 710032
Effect of miR-705 on osteogenic differentiation of mouse embryo osteoblast precursor cells MC3T3-E1
YANG Xiaohong1, YANG Kun2, LIAO Li3, JIN Yan3
1. Department of Prosthetics, School of Stomatology, Zunyi Medical College, Zunyi 563000, China;
2. Department of Periodontics, School of Stomatology, Zunyi Medical College, Zunyi 563000, China;
3. State Key Laboratory of Military Stomatology, Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi'an 710032, China
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摘要：

目的：观察微RNA（miR）-705对小鼠胚胎成骨细胞前体细胞（MC3T3-E1）成骨分化能力的影响。方法：将转染了miR-705模拟物、抑制物、对照物的MC3T3-E1细胞加入成骨诱导液，在其成骨诱导7 d时采用碱性磷酸酶（ALP）染色检测ALP的活性；在成骨诱导14 d时，用RT-PCR和蛋白质印迹技术检测Runt相关转录因子2（Runx2）和骨钙素（OCN）的mRNA及蛋白表达水平；在成骨诱导21 d时，以茜素红染色观察钙结节形成情况并作定量分析。结果：成骨诱导7 d时，模拟物组ALP活性降低，而抑制物组ALP活性升高（均P<0.05）；成骨诱导14 d时，模拟物组Runx2和OCN mRNA及蛋白表达水平较对照物组降低，而抑制物组则相反（均P<0.05）；成骨诱导21 d时，模拟物组钙结节最小，结节形成量下降，抑制物组钙结节较大且形成量较多（均P<0.05）。结论：miR-705对MC3T3-E1细胞成骨分化过程具有抑制作用。
关键词： 3T3细胞/药物作用;  成骨细胞/代谢;  微RNAs/治疗应用;  细胞转分化    
Abstract:

Objective: To investigate the effect of miR-705 on osteogenic differentiation of mouse embryo osteoblast precursor (MC3T3-E1) cells. Methods: miR-705 mimics, inhibitors and negative control were transfected into MC3T3-E1 cells. Alkaline phosphates (ALP) staining were performed and quantified after 7 days of osteogenic medium induction. The mRNA and protein expression levels of runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) were detected by real-time RT-PCR and Western blot after 14 days of osteogenic induction. Alizarin red staining was performed and quantified in MC3T3-E1 cells after 21 days of osteogenic induction. Results: After 7 days of osteogenic induction, ALP staining showed that overexpression of miR-705 significantly reduced ALP activity, whereas knockdown of miR-705 increased ALP activity (all P<0.05). Consistently, after 14 days of osteogenic induction, mRNA and protein expressions of Runx2 and OCN were suppressed by overexpression of miR-705, whereas they were promoted by knockdown of miR-705 (all P<0.05). After 21 days of osteogenic induction, alizarin red staining showed that overexpression of miR-705 significantly reduced the formation of mineralized node, the opposite results were found in miR-705 knockdown group (all P<0.05). Conclusion: miR-705 can inhibit the osteogenic differentiation of MC3T3-E1 cells.
Key words: 3T3 cells/drug effects    Osteoblasts/metabolism    MicroRNAs/therapeutic use    Cell transdifferentiation
收稿日期: 2016-07-05
CLC:  R34  
基金资助:

贵州省自然科学基金[黔科合（2013）2312]；遵义市红花岗基金[遵红科合社字（2014）07]
通讯作者: 金岩(1963-),男,博士,教授,博士生导师,主要从事骨、牙齿和神经的生长发育、愈合和修复中基因调控及分子信号传递机制研究;E-mail:yanjin@fumm.edu.cn;http://orcid.org/0000-0002-5952-9238     E-mail: yanjin@fumm.edu.cn
作者简介: 杨晓红(1976-),女,博士,副教授,主要从事微RNA与颌骨吸收的机制研究;E-mail:hxh2132000@163.com;http://orcid.org/0000-0003-3347-7553
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引用本文:

杨晓红 等. 微RNA-705对MC3T3-E1细胞成骨分化能力的影响[J]. 浙江大学学报（医学版）, 2016, 45(6): 575-580.

YANG Xiaohong, YANG Kun, LIAO Li, JIN Yan. Effect of miR-705 on osteogenic differentiation of mouse embryo osteoblast precursor cells MC3T3-E1. Journal of ZheJiang University(Medical Science), 2016, 45(6): 575-580.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2016.11.03    或    http://www.zjujournals.com/xueshu/med/CN/Y2016/V45/I6/575

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