Hey1表达对BMP-9诱导下C3H10T1/2细胞的成骨分化及增殖影响

发布时间：2021-02-27 19:36

　　目的探讨Notch信号通路重要靶点Hey1表达水平改变对BMP-9诱导C3H10T1/2细胞成骨分化与增殖的影响。方法构建过表达Hey1慢病毒LV-Hey1、抑制Hey1表达慢病毒LV-sh Hey1,分别感染C3H10T1/2细胞干预Hey1表达水平,以LV-Blank（空质粒）感染C3H10T1/2细胞作为对照;以荧光显微镜对慢病毒感染效果、实时荧光定量PCR以及Western blot对Hey1表达水平进行验证,筛选不同Hey1表达水平的稳定细胞系。用含BMP-9的条件培养基诱导不同Hey1表达水平的C3H10T1/2细胞（分别为BMP-9+C3H10T1/2组、BMP-9+C3H10T1/2-Hey1组、BMP-9+C3H10T1/2-sh Hey1组）,以正常培养基培养的细胞作为对照（C3H10T1/2组、C3H10T1/2-Blank组）。培养后48 h,实时荧光定量PCR及Western blot测定成骨分化相关转录因子Runx2、骨桥蛋白、骨钙素m RNA及蛋白表达水平;4、5、6、7 d行MTT检测及4、5、10 d行流式细胞仪测定细胞增殖能力;4、7 d时ELISA... 

【文章来源】：中国修复重建外科杂志. 2016,30(03)北大核心

【文章页数】：7 页

【部分图文】：

诱导培养4、7d各组ALP染色观察（×100）从左至右分别为C3H10T1/2组、C3H10T1/2-Blank组、BMP-9+C3H10T1/2组、BMP-9+C3H10T1/2-Hey1组、BMP-9+C3H10T1/2-shHey1组a培养4db培养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#▽*△统计值StatisticF=134.774，P=0.000F=123.249，P=0.000F=70.082，P=0.000#与C3H10T1/2组比较P<0.05，▽与C3H10T1/2-Blank组比较P<0.05，*与BMP-9+C3H10T1/2组比较P<0.05，△与BMP-9+C3H10T1/2-Hey1组比较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图7诱导培养4、7d各组ALP染色观察（×100）从左至右分别为C3H10T1/2组、C3H10T1/2-Blank组、BMP-9+C3H10T1/2组、BMP-9+C3H10T1/2-Hey1组、BMP-9+C3H10T1/2-shHey1组a培养4db培养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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