Upregulation of ETV2 Expression Promotes Endothelial Differentiation of Human Dental Pulp Stem Cells.
Upregulation of ETV2 Expression Promotes Endothelial Differentiation of Human Dental Pulp Stem Cells.
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ETV2表达的上调促进了人牙浆干细胞的内皮分化。
DOI：
10.1177/0963689720978739
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发表时间：
2021-01
影响因子：
3.3
通讯作者：
Xu J
中科院分区：
文献类型：
--
作者：
Li J;Zhu Y;Li N;Wu T;Zheng X;Heng BC;Zou D;Xu J
缺乏血管生成通常会阻碍新工程组织移植物在宿主体内的存活和整合。自体内皮细胞（ECs）是新生血管的理想细胞来源，但由于其稀缺性、缺乏增殖能力和分离后供体部位发病率的限制。本研究的目的是确定重组ETV2过表达是否能促进人牙髓干细胞（DPSCs）向内皮细胞的分化。从新鲜牙髓组织中提取DPSCs。通过慢病毒感染实现了ETV2在DPSCs中的过表达，并评估了细胞形态学变化。采用实时定量聚合酶链反应、western blot、免疫荧光染色、流式细胞术检测内皮特异性标志物mRNA和蛋白表达水平。在体外和体内分别采用成管实验和Matrigel塞实验来评估etv2转导细胞的血管生成潜能。此外，进行蛋白质组学分析，分析ETV2过表达后蛋白质表达的整体变化。慢病毒感染后，过表达etv2的DPSCs呈现内皮样形态。与对照DPSCs相比，在过表达etv2的DPSCs中，内皮特异性基因CD31、VE-Cadherin、VEGFR1和VEGFR2的mRNA和蛋白表达水平显著升高。此外，ETV2过表达增强了体外Matrigel上毛细血管样管的形成，以及体内新生血管的形成。此外，比较蛋白质组学分析显示，ETV2过表达上调血管内皮生长因子（VEGF）受体的表达，表明VEGF信号通路增加。综上所述，我们的研究结果表明，ETV2过表达显著增强了DPSCs的内皮分化。因此，本研究表明DPSCs可以成为组织工程应用的有前途的候选细胞来源。
The lack of vasculogenesis often hampers the survivability and integration of newly engineered tissue grafts within the host. Autologous endothelial cells (ECs) are an ideal cell source for neovascularization, but they are limited by their scarcity, lack of proliferative capacity, and donor site morbidity upon isolation. The objective of this study was to determine whether differentiation of human dental pulp stem cells (DPSCs) into the endothelial lineage can be enhanced by recombinant ETV2 overexpression. DPSCs were extracted from fresh dental pulp tissues. ETV2 overexpression in DPSCs was achieved by lentiviral infection and cellular morphological changes were evaluated. The mRNA and protein expression levels of endothelial-specific markers were assessed through quantitative real-time polymerase chain reaction, western blot, immunofluorescence staining, and flow cytometry. The tube formation assay and Matrigel plug assay were also performed to evaluate the angiogenic potential of the ETV2-transduced cells in vitro and in vivo, respectively. Additionally, proteomic analysis was performed to analyze global changes in protein expression following ETV2 overexpression. After lentiviral infection, ETV2-overexpressing DPSCs showed endothelial-like morphology. Compared with control DPSCs, significantly higher mRNA and protein expression levels of endothelial-specific genes, including CD31, VE-Cadherin, VEGFR1, and VEGFR2, were detected in ETV2-overexpressing DPSCs. Moreover, ETV2 overexpression enhanced capillary-like tube formation on Matrigel in vitro, as well as neovascularization in vivo. In addition, comparative proteomic profiling showed that ETV2 overexpression upregulated the expression of vascular endothelial growth factor (VEGF) receptors, which was indicative of increased VEGF signaling. Taken together, our results indicate that ETV2 overexpression significantly enhanced the endothelial differentiation of DPSCs. Thus, this study shows that DPSCs can be a promising candidate cell source for tissue engineering applications.
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影响因子：
20.1
作者：
Lee S;Park C;Han JW;Kim JY;Cho K;Kim EJ;Kim S;Lee SJ;Oh SY;Tanaka Y;Park IH;An HJ;Shin CM;Sharma S;Yoon YS
通讯作者：
Yoon YS
影响因子：
20.3
作者：
Otsu, Keishi;Das, Shonit;Bhattacharya, Jahar
通讯作者：
Bhattacharya, Jahar
影响因子：
3.7
作者：
Perugini V;Meikle ST;Guildford AL;Santin M
通讯作者：
Santin M
影响因子：
7.7
作者：
Liu, Fang;Li, Daofeng;Choi, Kyunghee
通讯作者：
Choi, Kyunghee
影响因子：
4.6
作者：
Wang, Ying;Kaiser, Mark S.;Essner, Jeffrey J.
通讯作者：
Essner, Jeffrey J.