NICD过表达对人牙髓干细胞细胞增殖及细胞迁移的影响

发布时间：2018-06-22 06:09

本文选题：牙髓干细胞 + NICD　；参考：《牙体牙髓牙周病学杂志》2014年04期

【摘要】：目的:利用Notch1的胞内段(Notch1 intracellular domain,NICD)基因过表达载体建立稳定过表达NICD的人牙髓干细胞(dental pulp stem cells,DPSCs)株,观察NICD过表达对DPSCs增殖、迁移能力的影响。方法:构建含NICD过表达的慢病毒颗粒,并将其转染至DPSCs构建过表达NICD的细胞株(DPSCs/NICD);以DPSCs/NICD为实验组,正常细胞株(DPSCs/wt)和空病毒转染的空载组(DPSCs/vector)作为对照,通过RT-PCR和Western Blot检测NICD mRNA及其蛋白的表达;细胞免疫荧光观察NICD在DPSCs上的表达位置;CCK8法检测细胞的增殖情况;流式细胞技术检测细胞周期中S期的改变;Transwll检测细胞的迁移能力。结果:与DPSCs/wt组和DPSCs/vector组相比,DPSCs/NICD组的NICDmRNA及其蛋白表达水平均明显增强(P0.05),表达位置在胞膜、胞浆及胞核;细胞的增殖速度加快、细胞周期中S期比例升高、迁移能力增强。结论:NICD过表达使DPSCs细胞的增殖和迁移能力增强。
[Abstract]:Aim: to study the effect of NICD overexpression on the proliferation and migration of human dental pulp stem cells (dental pulp stem cells) by using the over expression vector of Notch1 intracellular domain NICD gene. Methods: the lentivirus particles containing NICD overexpression were constructed and transfected into DPSCs to construct NICD overexpressed cell lines (DPSCs / NICD), DPSCsr / NICD was used as experimental group, normal cell line (DPSCsrwt) and empty virus transfected group (DPSCsP vector) were used as control group. The expression of NICD mRNA and its protein was detected by RT-PCR and Western blot, the expression of NICD in DPSCs was detected by cell immunofluorescence and the proliferation of DPSCs was detected by CCK8 method. The changes of S phase in cell cycle were detected by flow cytometry and the migration ability of cells was detected by Transwll. Results: compared with DPSCS / wt group and DPSCS / vector group, the expression level of NICD mRNA and protein in DPSCs / NICD group was significantly increased (P0.05), the expression location was in the cell membrane, cytoplasm and nucleus, the proliferation rate was accelerated, the S phase proportion of cell cycle was increased, and the migration ability was enhanced. Conclusion the proliferation and migration of DPSCs cells were enhanced by the overexpression of the cell line.
【作者单位】：第三军医大学新桥医院口腔科;
【基金】：国家自然科学基金资助项目(81170934)
【分类号】：R780.2

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