目的探討Notch信號通路重要靶點Hey1表達(dá)水平改變對BMP-9誘導(dǎo)C3H10T1/2細(xì)胞成骨分化與增殖的影響。方法構(gòu)建過表達(dá)Hey1慢病毒LV-Hey1、抑制Hey1表達(dá)慢病毒LV-sh Hey1,分別感染C3H10T1/2細(xì)胞干預(yù)Hey1表達(dá)水平,以LV-Blank（空質(zhì)粒）感染C3H10T1/2細(xì)胞作為對照;以熒光顯微鏡對慢病毒感染效果、實時熒光定量PCR以及Western blot對Hey1表達(dá)水平進(jìn)行驗證,篩選不同Hey1表達(dá)水平的穩(wěn)定細(xì)胞系。用含BMP-9的條件培養(yǎng)基誘導(dǎo)不同Hey1表達(dá)水平的C3H10T1/2細(xì)胞（分別為BMP-9+C3H10T1/2組、BMP-9+C3H10T1/2-Hey1組、BMP-9+C3H10T1/2-sh Hey1組）,以正常培養(yǎng)基培養(yǎng)的細(xì)胞作為對照（C3H10T1/2組、C3H10T1/2-Blank組）。培養(yǎng)后48 h,實時熒光定量PCR及Western blot測定成骨分化相關(guān)轉(zhuǎn)錄因子Runx2、骨橋蛋白、骨鈣素m RNA及蛋白表達(dá)水平;4、5、6、7 d行MTT檢測及4、5、10 d行流式細(xì)胞儀測定細(xì)胞增殖能力;4、7 d時ELISA... 

【文章來源】：中國修復(fù)重建外科雜志. 2016,30(03)北大核心

【文章頁數(shù)】：7 頁

【部分圖文】：

誘導(dǎo)培養(yǎng)4、7d各組ALP染色觀察（×100）從左至右分別為C3H10T1/2組、C3H10T1/2-Blank組、BMP-9+C3H10T1/2組、BMP-9+C3H10T1/2-Hey1組、BMP-9+C3H10T1/2-shHey1組a培養(yǎng)4db培養(yǎng)7dFig.7ALPstainingobservationofdierentgroupsat4and7days(×100)FromletorightforC3H10T1/2group,C3H10T1/2-Blankgroup,BMP-9+C3H10T1/2group,BMP-9+C3H10T1/2-Hey1group,andBMP-9+C3H10T1/2-shHey1group,respectivelyaAt4daysbAt7daysb

043#▽*BMP-9+C3H10T1/2-shHey10.165±0.003#▽*△0.571±0.093#▽*△0.215±0.025#▽*△統(tǒng)計值StatisticF=134.774，P=0.000F=123.249，P=0.000F=70.082，P=0.000#與C3H10T1/2組比較P<0.05，▽與C3H10T1/2-Blank組比較P<0.05，*與BMP-9+C3H10T1/2組比較P<0.05，△與BMP-9+C3H10T1/2-Hey1組比較P<0.05#ComparedwithC3H10T1/2group,P<0.05;▽comparedwithC3H10T1/2-Blankgroup,P<0.05;*comparedwithBMP-9+C3H10T1/2group,P<0.05;△comparedwithBMP-9+C3H10T1/2-Hey1group,P<0.05圖7誘導(dǎo)培養(yǎng)4、7d各組ALP染色觀察（×100）從左至右分別為C3H10T1/2組、C3H10T1/2-Blank組、BMP-9+C3H10T1/2組、BMP-9+C3H10T1/2-Hey1組、BMP-9+C3H10T1/2-shHey1組a培養(yǎng)4db培養(yǎng)7dFig.7ALPstainingobservationofdierentgroupsat4and7days(×100)FromletorightforC3H10T1/2group,C3H10T1/2-Blankgroup,BMP-9+C3H10T1/2group,BMP-9+C3H10T1/2-Hey1group,andBMP-9+C3H10T1/2-shHey1group,respectivelyaAt4daysbAt7daysab

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期刊論文
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