DLX5对牙源性间充质干细胞迁移及趋化作用的研究

doi:10.13591/j.cnki.kqyx.2022.05.003

摘要/Abstract

摘要： 目的探究distal-less homeobox 5 (DLX5)对牙源性间充质干细胞(dental tissue-derived mesenchymal stem cells, DT-MSCs)迁移及趋化的作用。方法利用逆转录病毒对根尖牙乳头干细胞(stem cells from apical papilla, SCAPs)进行转染,构建稳定过表达及敲减DLX5的细胞系。通过体外细胞划痕实验、transwell实验,探究DLX5对SCAPs迁移及趋化的影响。进一步观察DLX5过表达的SCAPs的条件培养基对牙周膜干细胞 (periodontal ligament stem cells, PDLSCs)迁移及趋化的影响。结果 ①逆转录病毒转染DLX5过表达的质粒后,DLX5在SCAPs中有效过表达;逆转录病毒转染DLX5敲减的质粒后,有效抑制了DLX5在SCAPs中的表达,与对照组相比,差异具有显著性。②过表达DLX5能明显促进SCAPs的迁移及趋化能力。③敲减DLX5能明显抑制SCAPs的迁移及趋化能力。④过表达DLX5的SCAPs的条件培养基促进了PDLSCs的迁移及趋化能力。结论 DLX5对SCAPs的迁移及趋化具有正向调控作用,过表达DLX5的SCAPs条件培养基可以促进PDLSCs的迁移及趋化能力。

关键词: distal-less homeobox 5, 牙源性间充质干细胞, 迁移, 趋化, 条件培养基

Abstract: Objective To reveal the function of Distal-Less Homeobox 5 (DLX5) in migration and chemotaxis of dental tissue-derived mesenchymal stem cells (DT-MSCs). Methods Retrovirus was used to transfect stem cells from apical papilla (SCAPs). After constructing the plasmid of DLX5, we overexpressed and knocked down the expression of DLX5. Real-time RT-PCR and Western Blot were used to detect the expression of DLX5. Scratch migration assays and transwell chemotaxis assays were used to examine the migration and chemotaxis capacity of SCAPs in vitro. The effects of DLX5-overexpressing conditioned medium on the migration and chemotaxis capacities of PDLSCs were tested by Scratch migration assays and transwell chemotaxis assays in vitro. Results ①After transferring the plasmid of DLX5 overexpression and knockdown with retrovirus, results of western blot and Real-time RT-PCR showed that the overexpression and knockdown efficiency of DLX5 was significantly high. ②The overexpression of DLX5 enhanced migration and chemotaxis capacities of SCAPs. ③The knockdown of DLX5 inhibited migration and chemotaxis capacities of SCAPs. ④The DLX5-overexpressing conditioned medium of SCAPs promoted migration and chemotaxis capacities of PDLSCs. Conclusion DLX5 significantly promotes migration and chemotaxis capacities of SCAPs. The DLX5-overexpressing conditioned medium of SCAPs promotes migration and chemotaxis capacities of PDLSCs.

Key words: distal-less homeobox 5, dental tissue-derived mesenchymal stem cell, migration, chemotaxis, conditioned medium

中图分类号:

R349.5

武涛涛, 张钰珩, 郑小雨, 刘畅, 马华瑞, 高润涛. DLX5对牙源性间充质干细胞迁移及趋化作用的研究[J]. 口腔医学, 2022, 42(5): 399-403.

WU Taotao, ZHANG Yuheng, ZHENG Xiaoyu, LIU Chang, MA Huarui, GAO Runtao. Effects of DLX5 on migration and chemotaxis of DT-MSCs[J]. Stomatology, 2022, 42(5): 399-403.

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