Hey1表達對BMP-9誘導(dǎo)下C3H10T1/2細胞的成骨分化及增殖影響
發(fā)布時間：2021-02-27 19:36
目的探討Notch信號通路重要靶點Hey1表達水平改變對BMP-9誘導(dǎo)C3H10T1/2細胞成骨分化與增殖的影響。方法構(gòu)建過表達Hey1慢病毒LV-Hey1、抑制Hey1表達慢病毒LV-sh Hey1,分別感染C3H10T1/2細胞干預(yù)Hey1表達水平,以LV-Blank（空質(zhì)粒）感染C3H10T1/2細胞作為對照;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表達水平進行驗證,篩選不同Hey1表達水平的穩(wěn)定細胞系。用含BMP-9的條件培養(yǎng)基誘導(dǎo)不同Hey1表達水平的C3H10T1/2細胞（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及蛋白表達水平;4、5、6、7 d行MTT檢測及4、5、10 d行流式細胞儀測定細胞增殖能力;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
【參考文獻】：
期刊論文
[1]Hey1基因參與調(diào)控BMP9誘導(dǎo)的C3H10T1/2細胞成骨分化[J]. 唐大剛,王淼,張艷亮,王孝林,李貴強,羅小輯. 中國生物工程雜志. 2015(06)
[2]攜帶siRbpj重組腺病毒載體的構(gòu)建及其對BMP9誘導(dǎo)骨髓間充質(zhì)干細胞成骨分化作用影響[J]. 張曉艷,楊倫韻,謝佳瑛,左國偉,羅進勇,唐敏. 重慶醫(yī)科大學(xué)學(xué)報. 2014(05)
[3]RUNX2對BMP9誘導(dǎo)的間充質(zhì)干細胞C3H10T1/2成骨分化的影響[J]. 趙艷芳,宋濤,劉躍亮,羅進勇. 中國生物工程雜志. 2013(02)
本文編號：3054703
【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
【參考文獻】：
期刊論文
[1]Hey1基因參與調(diào)控BMP9誘導(dǎo)的C3H10T1/2細胞成骨分化[J]. 唐大剛,王淼,張艷亮,王孝林,李貴強,羅小輯. 中國生物工程雜志. 2015(06)
[2]攜帶siRbpj重組腺病毒載體的構(gòu)建及其對BMP9誘導(dǎo)骨髓間充質(zhì)干細胞成骨分化作用影響[J]. 張曉艷,楊倫韻,謝佳瑛,左國偉,羅進勇,唐敏. 重慶醫(yī)科大學(xué)學(xué)報. 2014(05)
[3]RUNX2對BMP9誘導(dǎo)的間充質(zhì)干細胞C3H10T1/2成骨分化的影響[J]. 趙艷芳,宋濤,劉躍亮,羅進勇. 中國生物工程雜志. 2013(02)
本文編號：3054703
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