MicroRNA encoded by the HSV-1 latency-associated transcript anti-apoptotic function in human mesenchymal stem cells
MicroRNA encoded by the HSV-1 latency-associated transcript anti-apoptotic function in human mesenchymal stem cells
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HSV-1潜伏期相关转录物编码的MicroRNA在人间充质干细胞中的抗凋亡功能
DOI：
10.1007/s11434-008-0205-9
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发表时间：
2008-06
期刊：
影响因子：
--
通讯作者：
Li YongMei
中科院分区：
文献类型：
--
作者：
Li WeiXia;Du WenCai;Zhu Ze;Zhang Jing;Li GuangMing;Li YongMei
人类间充质干细胞（HMSC）已成为一种有前途的细胞类型，用于组织再生和用作离体基因递送载体。单纯疱疹病毒-1（HSV-1）具有许多载体特征，这使其特别适合于HMSC应用。本研究利用特异性茎环反向引物进行实时荧光定量PCR检测HMSCs细胞质中HSV-1潜伏相关转录本（microRNA-LAT）编码的成熟microRNA水平。针对microRNA-LAT、TGF-β1和SMAD 3化学合成三种小干扰RNA（siRNA）。结果表明，HSV-1和microRNA-LAT阻止HMSC经历顺铂诱导的凋亡。与仅用顺铂处理的细胞相比，用HSV-1和microRNA-LAT处理的细胞凋亡现象显著减少。流式细胞仪检测两组细胞凋亡率均较低（P<0.05）。RT-PCR和western blotting分析结果证实，HSV-1和microRNA-LAT处理后，TGF-β1和SMAD 3的mRNA和蛋白水平均显著降低。通过这些方法敲除整合的TGF-β1信号通路组分。此外，顺铂处理的HMSCs中成熟microRNA-LAT的水平降低。总之，HSV-1和microRNA-LAT通过下调TGF-β1信号通路对HMSC发挥其抗凋亡作用。因此，具有抗凋亡作用的HSV-1在其microRNA-LAT中天然编码，是开发用于HMSC载体的良好候选者。
Human mesenchymal stem cells (HMSCs) have emerged as a promising cell type for tissue repopulating and for use as ex vivo gene delivery vehicles. Herpes simplex virus-1 (HSV-1) possesses many vector features, which make it especially suitable for HMSC applications. Here we performed real-time RT-PCR to detect the level of mature microRNA encoded by the HSV-1 latency-associated transcript (microRNA-LAT) in HMSCs cytoplasm with a specific stem loop reverse primer. Three small interfering RNAs (siRNAs) were chemically synthesized towards microRNA-LAT, TGF-β1 and SMAD3. The results demonstrate that HSV-1 and microRNA-LAT prevented HMSCs from undergoing cisplatin-induced apoptosis. In comparison with cells only treated with cisplatin, the apoptosis phenomenon with HSV-1 and microRNA-LAT were markedly reduced. The apoptosis rates of these two groups were both lower (P<0.05) as determined by flow cytometry analysis. The results of RT-PCR and western blotting analysis confirmed that the mRNA and protein levels of TGF-β1 and SMAD3 were significantly decreased with treatment of HSV-1 and microRNA-LAT. Integral TGF-β1 signalling pathway components were by these means knocked down. Moreover, the levels of the mature microRNA-LAT were decreased in cisplatin-treated HMSCs. In conclusion, HSV-1 and microRNA-LAT exert their anti-apoptotic effect on HMSCs by down-regulation of the TGF-β1 signalling pathway. Thus HSV-1, having anti-apoptotic effect naturally encoded in its microRNA-LAT, is a good candidate to be developed for HMSC vector.
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DOI：
10.1007/978-0-387-40049-5_29
发表时间：
2006
期刊：
--
影响因子：
--
作者：
Yan Zeng;B. Cullen
通讯作者：
Yan Zeng;B. Cullen
影响因子：
--
作者：
Zhuozhuang Lu;Zu-ze Wu;Qun-wei Zhang;Hua Wang;X. Jia;H. Duan;Li-Sheng Wang
通讯作者：
Zhuozhuang Lu;Zu-ze Wu;Qun-wei Zhang;Hua Wang;X. Jia;H. Duan;Li-Sheng Wang
影响因子：
--
作者：
Haiyun Pei;Yunfang Wang;Xuetao Pei
通讯作者：
Haiyun Pei;Yunfang Wang;Xuetao Pei
影响因子：
5.4
作者：
Inman, M;Perng, GC;Jones, C
通讯作者：
Jones, C
影响因子：
9.8
作者：
John B;Enright AJ;Aravin A;Tuschl T;Sander C;Marks DS
通讯作者：
Marks DS