Lnc-NTF3-5对上颌窦黏膜干细胞成骨分化的作用

The effect of lnc-NTF3-5 on osteogenic differentiation of maxillary sinus membrane stem cells in vitro

目的:探讨lnc-NTF3-5对上颌窦黏膜干细胞成骨分化的作用。方法:体外培养上颌窦黏膜干细胞(maxillary sinus mucosa stem cells,MSMSCs),经成骨诱导培养后,利用荧光定量PCR(RT-PCR)检测lnc-NTF-5的表达。使lnc-NTF-5过表达或沉默表达,RT-PCR检测Runt相关转录因子2(runt-related transcription factor 2,RUNX2)、骨钙蛋白(osteocalcin,OCN)和碱性磷酸酶(alkaline phosphatase,ALP)m RNA的表达,Western Blot检测RUNX2、OCN和ALP蛋白的表达,茜素红染色观察钙结节形成情况。结果:Lnc-NTF3-5在MSMSCs成骨分化过程中表达量升高(P<0.001)。MSMSCs过表达lnc-NTF3-5后,成骨相关标志物RUNX2、OCN和ALP的m RNA和蛋白表达水平明显升高(P<0.05),茜素红染色钙结节数目增多;相反,沉默lnc-NTF3-5的MSMSCs中RUNX2、OCN、ALP m RNA和蛋白表达水平明显降低(P<0.01),茜素红染色钙结节数目减少。结论:Lnc-NTF3-5促进MSMSCs的成骨分化。

Objective：To investigate the effect of lnc-NTF-5 on osteogenic differentiation of maxillary sinus mucosa stem cells. Methods： Maxillary sinus mucosa stem cells（ MSMSCs） were cultured in vitro. After osteogenic induction,the expression of lnc-NTF-5 was detected by RT-PCR. RT-PCR was used to detect the expression of runt-related transcription factor 2（ RUNX2）,osteocalcin（ OCN） and alkaline phosphatase（ ALP） m RNA,and the expression of RUNX2,OCN and ALP proteins was detected by Western Blot,and alizarin red staining was used to observe the formation of calcium nodules after transfected with lnc-NTF3-5 overexpression plasmid or sh-lnc-NTF3-5. Results： The expression of lnc-NTF3-5 increased during osteogenic differentiation of MSMSCs（ P〈0. 001）. In lnc-NTF3-5-overexpressed-MSMSCs,the m RNA and protein level of osteogenic markers RUNX2,OCN and ALP were significantly higher（ P〈0. 05）,and the number of calcium nodules increased. On the contrary,in the lnc-NTF3-silencing MSMSCs,the expression of RUNX2,OCN,ALP on m RNA and protein level was significantly lower（ P〈0. 01）,and the number of calcium nodules decreased. Conclusion： Lnc-NTF3-5 can promote osteogenic differentiation of maxillary sinus mucosa stem cells.