Cardiomyocyte protection by GATA-4 gene engineered mesenchymal stem cells is partially mediated by translocation of miR-221 in microvesicles.
Cardiomyocyte protection by GATA-4 gene engineered mesenchymal stem cells is partially mediated by translocation of miR-221 in microvesicles.
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DOI：
10.1371/journal.pone.0073304
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Xu M
中科院分区：
文献类型：
--
作者：
Yu B;Gong M;Wang Y;Millard RW;Pasha Z;Yang Y;Ashraf M;Xu M
microRNA（miRs）是一类新型的非编码小RNA，参与细胞增殖、分化、发育和死亡。在这项研究中，我们发现miR-221通过微囊泡（MV）的易位在由加塔-4过表达的间充质干细胞（MSC）介导的心脏保护中起重要作用。收获成年大鼠骨髓MSC和新生大鼠心室心肌细胞（CM）作为原代培养物。使用鼠干细胞病毒（pMSCV）逆转录病毒表达系统用加塔-4（MSC加塔-4）转导MSC。空载体转染用作对照（MSCO 2）。通过实时PCR评估miR的表达，并使用原位杂交（ISH）进行定位。从MSC培养物中收集的MV的特征是CD 9、CD 63和HSP 70的表达，并用电子显微镜拍照。通过乳酸脱氢酶（LDH）释放、CM摄取MTS和caspase 3/7活性评价MSC培养物条件培养基（CdM）在缺氧期间提供的细胞保护作用。miR-221/222在MSC中的表达显著高于CM，并且miR-221在MSCGATA-4中上调。MSC过表达miR-221通过减少p53上调的凋亡调节因子（p53 modulator of apoptosis，p53）的表达显著增强了心脏保护作用。此外，CM与MSC共培养后，CD 45 A的表达显著降低。来源于MSC的MV表达高水平的miR-221，并且通过CM快速内化，如从Time-Lapse Imaging System获得的图像中所记录的。我们的研究结果表明，MSCGATA-4的心脏保护作用可能部分受到MV中所含抗凋亡miR的转移的调节。
microRNAs (miRs), a novel class of small non-coding RNAs, are involved in cell proliferation, differentiation, development, and death. In this study, we found that miR-221 translocation by microvesicles (MVs) plays an important role in cardioprotection mediated by GATA-4 overexpressed mesenchymal stem cells (MSC). Adult rat bone marrow MSC and neonatal rat ventricle cardiomyocytes (CM) were harvested as primary cultures. MSC were transduced with GATA-4 (MSCGATA-4) using the murine stem cell virus (pMSCV) retroviral expression system. Empty vector transfection was used as a control (MSCNull). The expression of miRs was assessed by real-time PCR and localized using in situ hybridization (ISH). MVs collected from MSC cultures were characterized by expression of CD9, CD63, and HSP70, and photographed with electron microscopy. Cardioprotection during hypoxia afforded by conditioned medium (CdM) from MSC cultures was evaluated by lactate dehydrogenase (LDH) release, MTS uptake by CM, and caspase 3/7 activity. Expression of miR-221/222 was significantly higher in MSC than in CM and miR-221 was upregulated in MSCGATA-4. MSC overexpression of miR-221 significantly enhanced cardioprotection by reducing the expression of p53 upregulated modulator of apoptosis (PUMA). Moreover, expression of PUMA was significantly decreased in CM co-cultured with MSC. MVs derived from MSC expressed high levels of miR-221, and were internalized quickly by CM as documented in images obtained from a Time-Lapse Imaging System. Our results demonstrate that cardioprotection by MSCGATA-4 may be regulated in part by a transfer of anti-apoptotic miRs contained within MVs.
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影响因子：
3.7
作者：
Boomsma RA;Geenen DL
通讯作者：
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影响因子：
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