过表达Notch1的胞内段基因对人牙周膜干细胞增殖能力影响

doi:10.13701/j.cnki.kqyxyj.2019.01.007

口腔医学研究 ›› 2019, Vol. 35 ›› Issue (1): 28-32.DOI: 10.13701/j.cnki.kqyxyj.2019.01.007

过表达Notch1的胞内段基因对人牙周膜干细胞增殖能力影响

龙晏, 邱申彩, 李淑慧, 舍玉秀, 王娜, 陈晓燕, 吴佩玲^*

新疆医科大学第二附属医院口腔科新疆乌鲁木齐 830063

收稿日期:2018-07-09 出版日期:2019-01-18 发布日期:2019-01-28
通讯作者: 吴佩玲,E-mail:wplkq@sina.com
作者简介:龙晏(1993~ ),女,重庆永川人,硕士在读,住院医师,主要从事牙体牙髓病的临床研究工作。
基金资助:
国家自然科学基金(编号:81460103)

Effect of Over Expression of NICD Gene on Proliferation of Human Periodontal Ligament Stem Cells

LONG Yan, QIU Shen-cai, LI Shu-hui, SHE Yu-xiu, WANG Na, CHEN Xiao-yan, WU Pei-ling^*

Department of Stomatology, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830063, China

Received:2018-07-09 Online:2019-01-18 Published:2019-01-28

摘要/Abstract

摘要： 目的:探讨过表达Notch1的胞内段(Notch1 intracellular domain,NICD)基因对人牙周膜干细胞(periodontal ligament stem cells,PDLSCs)增殖能力的影响。方法:构建含NICD基因过表达的逆转录病毒颗粒,并将其转染至牙周膜干细胞,构建过表达NICD的细胞株PDLSCs/NICD,转染48 h后观察细胞一般状态。PDLSCs/NICD为实验组,以PDLSCs/wt为正常细胞组和PDLSCs/vector空病毒转染组作为对照,通过实时定量聚合酶链锁反应(quantitative Real-time-Polymerase chain reaction,qRT-PCR)和Western blot检测NICD mRNA及其蛋白的表达; 细胞计数试剂盒(cell counting kit-8,CCK-8)法检测细胞的增殖情况;Transwell实验检测细胞的迁移能力。结果:Western blot和qRT-PCR结果显示,与PDLSCs/wt组和PDLSCs/vector组比较,PDLSCs/NICD组的NICD蛋白表达水平及其mRNA表达水平均明显升高(P＜0.05);CCK-8结果提示,与PDLSCs/vector组比较,PDLSCs/NICD组细胞增殖速度加快,Transwell实验结果显示,PDLSCs/NICD组的细胞迁移(40.20±1.74)明显高于PDLSCs/vector组(21.20±1.18)。结论:NICD基因过表达的人PDLSCs在一定程度上增殖及迁移能力加强。

关键词: 牙周膜干细胞, Notch1的胞内段, 转染, 增殖, 迁移

Abstract: Objective: To investigate the effect of Notch1 intracellular domain (NICD) gene on the proliferation of human periodontal ligament stem cells (PDLSCs). Methods: The retroviral particles with over expression of Notch1 (NICD) gene were constructed and transfected into the PDLSCs. The general state of the cells was observed after 48h transfection. PDLSCs/NICD was set as the experimental group, PDLSCs/wt was set as normal group, and PDLSCs/empty vector was set as control group. The expressions of NICD mRNA and protein were detected by Real-time PCR and Western Blot, the proliferation of cells was detected by CCK8 method, and the migration ability of the cells was detected. Results: The expression levels of protein and NICD mRNA in the PDLSCs/NICD group were obviously enhanced in contrast to PDLSCs/wt group and the PDLSCs/empty vector group. Cell migration of PDLSCs/NICD group (40.20±1.738) was significantly higher than that of PDLSCs/vector group (21.20±1.182). Conclusion: Over expression of NICD enhanced the proliferation and migration ability of human PDLSCs to a certain extent.

Key words: Periodontal ligament stem cells, NICD, Transfection, Proliferation, Migration

龙晏, 邱申彩, 李淑慧, 舍玉秀, 王娜, 陈晓燕, 吴佩玲. 过表达Notch1的胞内段基因对人牙周膜干细胞增殖能力影响[J]. 口腔医学研究, 2019, 35(1): 28-32.

LONG Yan, QIU Shen-cai, LI Shu-hui, SHE Yu-xiu, WANG Na, CHEN Xiao-yan, WU Pei-ling. Effect of Over Expression of NICD Gene on Proliferation of Human Periodontal Ligament Stem Cells[J]. Journal of Oral Science Research, 2019, 35(1): 28-32.

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过表达Notch1的胞内段基因对人牙周膜干细胞增殖能力影响

Effect of Over Expression of NICD Gene on Proliferation of Human Periodontal Ligament Stem Cells

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