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Title of Journal: Chin Sci Bull

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Abbravation: Chinese Science Bulletin

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SP Science in China Press

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DOI

10.1002/tera.1420360323

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1861-9541

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MicroRNA encoded by the HSV1 latencyassociated t

Authors: Zhang Jing Li YongMei Du WenCai Li GuangMing Li WeiXia Zhu Ze
Publish Date: 2008/06/05
Volume: 53, Issue: 11, Pages: 1691-1698
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Abstract

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 virus1 HSV1 possesses many vector features which make it especially suitable for HMSC applications Here we performed realtime RTPCR to detect the level of mature microRNA encoded by the HSV1 latencyassociated transcript microRNALAT in HMSCs cytoplasm with a specific stem loop reverse primer Three small interfering RNAs siRNAs were chemically synthesized towards microRNALAT TGFβ1 and SMAD3 The results demonstrate that HSV1 and microRNALAT prevented HMSCs from undergoing cisplatininduced apoptosis In comparison with cells only treated with cisplatin the apoptosis phenomenon with HSV1 and microRNALAT were markedly reduced The apoptosis rates of these two groups were both lower P005 as determined by flow cytometry analysis The results of RTPCR and western blotting analysis confirmed that the mRNA and protein levels of TGFβ1 and SMAD3 were significantly decreased with treatment of HSV1 and microRNALAT Integral TGFβ1 signalling pathway components were by these means knocked down Moreover the levels of the mature microRNALAT were decreased in cisplatintreated HMSCs In conclusion HSV1 and microRNALAT exert their antiapoptotic effect on HMSCs by downregulation of the TGFβ1 signalling pathway Thus HSV1 having antiapoptotic effect naturally encoded in its microRNALAT is a good candidate to be developed for HMSC vectorSupported by the National Natural Science Foundation of China Grant No 30772483 Scientific Research Foundation for the Returned Overseas Chinese Scholars State Education Ministry of P R China and the Tianjin Municipal Science and Technology Committee Grant No 05YFJMJC03900

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