Epigenetic regulation of connective tissue growth factor by MicroRNA-214 delivery in exosomes from mouse or human hepatic stellate cells.
Epigenetic regulation of connective tissue growth factor by MicroRNA-214 delivery in exosomes from mouse or human hepatic stellate cells.
复制标题
DOI：
10.1002/hep.26768
复制
发表时间：
2014-03
期刊：
影响因子：
13.5
通讯作者：
Brigstock, David R.
中科院分区：
文献类型：
--
作者：
Chen, Li;Charrier, Alyssa;Zhou, Yu;Chen, Ruju;Yu, Bo;Agarwal, Kitty;Tsukamoto, Hidekazu;Lee, L. James;Paulaitis, Michael E.;Brigstock, David R.
结缔组织生长因子(CCN2)促进肝星状细胞(HSC)的纤维化形成。在这里，我们表明，在纤维化或脂肪变性的肝脏，或在培养激活或乙醇处理的原代小鼠HSC中，CCN2的上调与microRNA-214(miR-214)的相互下调有关。通过保护子或报告分子实验研究CCN2的3‘非翻译区，我们发现肝纤维化诱导的HSC中CCN2的表达是由于miR-214的表达减少，而miR-214则通过直接与CCN2的3’-非翻译区结合来抑制CCN2的表达。此外，miR214存在于肝星状细胞外切体中，为双膜囊泡，直径50-150 nm，带负电荷(−为26 mV)，CD9阳性。用外切体抑制剂GW4869预处理细胞，可降低外切体中的MIR-214水平，但不能降低细胞裂解产物中的MIR-214水平。将miR-214基因修饰的供体HSC与CCN2 3‘-UTR荧光素酶报告基因修饰的受体HSC共培养，发现野生型CCN2 3’-UTR报告基因对miR-214和外切体的依赖性调节作用，而突变的CCN23‘-UTR报告基因表达缺失miR-214结合位点。HSC的外切体是小鼠原代肝细胞摄取miR-214的通道。MiR-214下调CCN2的表达也发生在人LX-2 HSC中，这与人CCN23 3‘-UTR中保守的miR-214结合位点一致。在LX-2细胞中，MIR-214通过外切体被运送到受体LX-2细胞或人肝癌细胞中，导致CCN2 3‘-非编码区活性或CCN2下游靶点的表达受到抑制，包括αSMA或胶原。小鼠实验性纤维化与循环miR-214水平降低有关。MiR-214的外体转移是一种调控CCN2依赖的纤维化形成的范例，并将纤维化途径确定为外体miR的表观遗传调控的靶点。
Connective tissue growth factor (CCN2) drives fibrogenesis in hepatic stellate cells (HSC). Here we show that CCN2 up-regulation in fibrotic or steatotic livers, or in culture-activated or ethanol-treated primary mouse HSC is associated with a reciprocal down-regulation of microRNA-214 (miR-214). By using protector or reporter assays to investigate the 3′-untranslated region (UTR) of CCN2 mRNA, we found that induction of CCN2 expression in HSC by fibrosis-inducing stimuli was due to reduced expression of miR-214 which otherwise inhibited CCN2 expression by directly binding to the CCN2 3′-UTR. Additionally, miR-214 was present in HSC exosomes, which were bi-membrane vesicles, 50–150nm in diameter, negatively charged (−26mV), and positive for CD9. MiR-214 levels in exosomes but not in cell lysates were reduced by pre-treatment of the cells with the exosome inhibitor, GW4869. Co-culture of miR-214-transfected donor HSC with CCN2 3′-UTR luciferase reporter-transfected recipient HSC resulted in miR-214- and exosome-dependent regulation of a wild type CCN2 3′-UTR reporter but not of a mutant CCN2 3′-UTR reporter lacking the miR-214 binding site. Exosomes from HSC were a conduit for uptake of miR-214 by primary mouse hepatocytes. Down-regulation of CCN2 expression by miR-214 also occurred in human LX-2 HSC, consistent with a conserved miR-214 binding site in the human CCN2 3′-UTR. MiR-214 in LX-2 cells was shuttled via exosomes to recipient LX-2 cells or human HepG2 hepatocytes, resulting in suppression of CCN2 3′-UTR activity or expression of CCN2 downstream targets, including αSMA or collagen. Experimental fibrosis in mice was associated with reduced circulating miR-214 levels. Exosomal transfer of miR-214 is a paradigm for the regulation of CCN2-dependent fibrogenesis and identifies fibrotic pathways as targets of epigenetic regulation by exosomal miRs.
登录
查看更多内容
影响因子：
4
作者：
Noetel A;Kwiecinski M;Elfimova N;Huang J;Odenthal M
通讯作者：
Odenthal M
影响因子：
24.5
作者：
Pan, Qiuwei;Ramakrishnaiah, Vedashree;van der Laan, Luc J. W.
通讯作者：
van der Laan, Luc J. W.
影响因子：
13.5
作者：
Roderburg, Christoph;Urban, Gerd-Willem;Luedde, Tom
通讯作者：
Luedde, Tom
影响因子：
3.9
作者：
Ghiassi-Nejad Z;Friedman SL
通讯作者：
Friedman SL
细胞间纳米层介导的microRNA转移：肝癌细胞生长的环境调节机制。
影响因子：
13.5
作者：
Kogure, Takayuki;Lin, Wen-Lang;Yan, Irene K.;Braconi, Chiara;Patel, Tushar
通讯作者：
Patel, Tushar