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机构地区:[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
出 处:《浙江大学学报(医学版)》2016年第6期575-580,共6页Journal of Zhejiang University(Medical Sciences)
基 金: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]
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7d时采用碱性磷酸酶(ALP)染色检测ALP的活性;在成骨诱导14d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21d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7d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14d时,模拟物组Runx2和OCNmRNA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21d时,模拟物组钙结节最小,结节形成量下降,抑制物纽钙结节较大且形成量较多(均P〈0.05)。结论:miR-705对Mc313-E1细胞成骨分化过程具有抑制作用。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 osteogenie 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.
关 键 词:3T3细肜药物作用 成骨细胞/代谢 微RNAs/治疗应用 细胞转分化
分 类 号:R329.2[医药卫生—人体解剖和组织胚胎学]
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