Lentiviral vector-mediated co-overexpression of VEGF and Bcl-2 improves mesenchymal stem cell survival and enhances paracrine effects in vitro.
Lentiviral vector-mediated co-overexpression of VEGF and Bcl-2 improves mesenchymal stem cell survival and enhances paracrine effects in vitro.
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DOI：
10.3892/ijmm.2017.3019
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发表时间：
2017-08
影响因子：
5.4
通讯作者：
Chen M
中科院分区：
文献类型：
--
作者：
Ni X;Ou C;Guo J;Liu B;Zhang J;Wu Z;Li H;Chen M
间充质干细胞（MSC）移植已成为缺血性心脏病的一种有前途的治疗方法，然而，移植细胞的低存活率限制了其治疗效果。本研究的目的是调查是否双重基因修饰的血管内皮生长因子（VEGF）和B细胞淋巴瘤-2（Bcl-2）赋予更高的表达水平的目标基因，更好的生存和更强的旁分泌效应在骨髓间充质干细胞在不利的环境比修改的个别基因。为此，通过使用自切割T2 A肽序列连接并实现VEGF和Bcl-2的共过表达来构建慢病毒载体。转染大鼠MSC以获得表现出稳定过表达的细胞系。采用体外氧糖剥夺（OGD）模型模拟缺血微环境，观察细胞凋亡、自噬和旁分泌效应。与单个基因修饰的MSCs和对照MSCs相比，双重基因修饰的MSCs具有更高的目的基因表达水平、更快的增殖、更少的凋亡、更少的自噬和更强的旁分泌效应。此外，在体外OGD模型中，发现自噬的抑制有助于细胞凋亡的抑制。总体而言，这些数据表明VEGF和Bcl-2的共过表达通过抑制凋亡、抑制自噬和增强旁分泌效应来保护缺血环境中的MSC。
Mesenchymal stem cell (MSC) transplantation has emerged as a promising therapy for ischemic heart disease; however, the low survival rate of transplanted cells limits their therapeutic efficacy. The aim of this study was to investigate whether the dual genetic modification of vascular endothelial growth factor (VEGF) and B-cell lymphoma-2 (Bcl-2) confers a higher expression level of the target genes, better survival and a stronger paracrine effect in MSCs in an adverse environment than the modification of the individual genes. For this purpse, a lentiviral vector was constructed by using a self-cleaving T2A peptide sequence to link and achieve the co-overexpression of VEGF and Bcl-2. Rat MSCs were transfected to obtain cell lines that exhibited a stable overexpression. An in vitro model of oxygen glucose deprivation (OGD) was applied to mimic the ischemic microenvironment, and cell apoptosis, autophagy and the paracrine effects were then determined. Compared with the MSCs in which individual genes were modified and the control MSCs, the MSCs which were subjected to dual genetic modification had a higher expression level of the target genes, a more rapid proliferation, reduced apoptosis, decreased autophagy and an enhanced paracrine effect. Furthermore, the suppression of autophagy was found to contribute to the inhibition of apoptosis in this in vitro OGD model. On the whole, these data indicate that the co-overexpression of VEGF and Bcl-2 protects MSCs in an ischemic environment by inhibiting apoptosis, suppressing autophagy and enhancing the paracrine effects.
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影响因子：
4.3
作者：
Wang X;Zhang J;Zhang F;Li J;Li Y;Tan Z;Hu J;Qi Y;Li Q;Yan B
通讯作者：
Yan B
影响因子：
--
作者：
Lee S;Choi E;Cha MJ;Hwang KC
通讯作者：
Hwang KC
影响因子：
20.1
作者：
Duran JM;Makarewich CA;Sharp TE;Starosta T;Zhu F;Hoffman NE;Chiba Y;Madesh M;Berretta RM;Kubo H;Houser SR
通讯作者：
Houser SR
影响因子：
5.2
作者：
Wang, Xiaohong;Zhao, Tiemin;Huang, Wei;Wang, Tao;Qian, Jiang;Xu, Meifeng;Kranias, Evangelia G.;Wang, Yigang;Fan, Guo-Chang
通讯作者：
Fan, Guo-Chang
VEGF 和 Bcl-2 在神经母细胞瘤缺氧反应中通过 MAPKs 信号通路相互作用
影响因子：
4
作者：
Wang, Duoduo;Weng, Qinjie;Yang, Bo
通讯作者：
Yang, Bo