目的:探討miR-21與BMP9之間的關(guān)系,明確miR-21在BMP9誘導(dǎo)間充質(zhì)干細(xì)胞成骨分化中的作用。方法:（1）Ad-BMP9感染C3H10T1/2細(xì)胞,Real-time-PCR檢測miR-21表達(dá)。RTPCR檢測ALP的表達(dá)。（2）MiR-21轉(zhuǎn)染C3H10T1/2細(xì)胞,Real-time-PCR檢測miR-21和BMP9表達(dá)。（3）MiR-21和BMP9-CM處理C3H10 T1/2細(xì)胞,ALP活性和染色實(shí)驗(yàn)檢測C3H10 T1/2細(xì)胞早期成骨能力。茜素紅S染色實(shí)驗(yàn)檢測鈣鹽沉積情況。（4）MiR-21和BMP9-CM處理C3H10T1/2細(xì)胞,Real-time-PCR檢測成骨分化相關(guān)因子ALP,OCN的表達(dá)。（5）MiR-21和BMP9-CM處理C3H10T1/2細(xì)胞,Western blot檢測p-Smad1/5蛋白水平的表達(dá)。結(jié)果:（1）BMP9暫時(shí)降低miR-21的表達(dá)。MiR-21也可以暫時(shí)降低BMP9的表達(dá)。（2）MiR-21可以協(xié)同BMP9增強(qiáng)ALP和鈣鹽沉積。（3）MiR-21協(xié)同BMP9增加了p-Smad1/5蛋白水平的表達(dá)。結(jié)論:MiR-21與BMP9存在... 

【文章來源】：中國生物工程雜志. 2016,36(02)北大核心CSCD

【文章頁數(shù)】：8 頁

【部分圖文】：

miＲ-21在BMP9誘導(dǎo)C3H10T1/2細(xì)胞成骨分化中的表達(dá)Fig．1ThetemporarydownregulationofmiＲ-21duringBMP9-inducedosteogenicdifferentiationofC3H10T1/2cell(a)Ad-BMP9andAd-GFPcaninfectC3H10T1/2cell(×100)(b)ExpressionofALPdetectedbyＲT-PCＲ(c)ExpressionofmiＲ-21＊＊＊＊＊

2016，36(2)宋琪玲等:MiＲ-21協(xié)同BMP9促進(jìn)間充質(zhì)干細(xì)胞C3H10T1/2成骨分化圖2過表達(dá)miＲ-21，BMP9在C3H10T1/2細(xì)胞中的表達(dá)變化Fig．2ThetemporarydownregulationofBMP9mＲNAbytransferringmiＲ-21intoC3H10T1/2cell(a)ExpressionofmiＲ-21detectedbyＲeal-timePCＲ＊＊＊P＜0．001comparedwithNCgroup(b)ExpressionofBMP9at1daydetectedbyＲeal-timePCＲ＊＊P＜0．01comparedwithNCgroup(c)ExpressionofBMP9at5daydetectedbyＲeal-timePCＲ*P＜0．5comparedwithNCgroup圖3miＲ-21促進(jìn)了BMP9誘導(dǎo)的C3H10T1/2細(xì)胞早期成骨分化Fig．3OverexpressionmiＲ-21enhancesBMP9-inducedearlyosteogenicdifferentiantionofC3H10T1/2cell(a)ALPstainingresultswererecordedinhighermagnifications(×100)(b)ALPstainingresultswererecordedinlowermagnifications(c)ALPactivitydeterminedbyALPquantitativeassay*P＜0．5comparedwithNC+BMP9group;＊＊P＜0．01comparedwithmiＲ-21group25

2016，36(2)宋琪玲等:MiＲ-21協(xié)同BMP9促進(jìn)間充質(zhì)干細(xì)胞C3H10T1/2成骨分化圖2過表達(dá)miＲ-21，BMP9在C3H10T1/2細(xì)胞中的表達(dá)變化Fig．2ThetemporarydownregulationofBMP9mＲNAbytransferringmiＲ-21intoC3H10T1/2cell(a)ExpressionofmiＲ-21detectedbyＲeal-timePCＲ＊＊＊P＜0．001comparedwithNCgroup(b)ExpressionofBMP9at1daydetectedbyＲeal-timePCＲ＊＊P＜0．01comparedwithNCgroup(c)ExpressionofBMP9at5daydetectedbyＲeal-timePCＲ*P＜0．5comparedwithNCgroup圖3miＲ-21促進(jìn)了BMP9誘導(dǎo)的C3H10T1/2細(xì)胞早期成骨分化Fig．3OverexpressionmiＲ-21enhancesBMP9-inducedearlyosteogenicdifferentiantionofC3H10T1/2cell(a)ALPstainingresultswererecordedinhighermagnifications(×100)(b)ALPstainingresultswererecordedinlowermagnifications(c)ALPactivitydeterminedbyALPquantitativeassay*P＜0．5comparedwithNC+BMP9group;＊＊P＜0．01comparedwithmiＲ-21group25

【參考文獻(xiàn)】：
期刊論文
[1]TGF-β/BMP signaling and other molecular events: regulation of osteoblastogenesis and bone formation[J]. Md Shaifur Rahman,Naznin Akhtar,Hossen Mohammad Jamil,Rajat Suvra Banik,Sikder M Asaduzzaman.  Bone Research. 2015(01)
[2]Spry1在miR-21調(diào)控人骨髓間充質(zhì)干細(xì)胞成骨分化中的作用[J]. 楊楠,周威,王光,丁寅,金巖.  中國組織工程研究. 2014(32)
[3]miR-30a抑制BMP9誘導(dǎo)間充質(zhì)干細(xì)胞C3H10T1/2的成骨分化[J]. 李寶林,白慧麗,張汝益,嚴(yán)樹涓,何方,楊丹丹,劉晨,施瓊.  中國生物工程雜志. 2013(11)



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