MicroRNAs regulate self-renewal and differentiation of mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2012.49.031

Abstract

Abstract:

BACKGROUND: MicroRNAs (miRNA) play an important role in self-renewal and differentiation of mesenchymal stem cells by modulating expression of some key transcription factors, receptors, and some specific proteins related to the developmental process.
OBJECTIVE: To discuss the regulatory roles of miRNAs in self-renewal and differentiation of mesenchymal stem cells.
METHODS: PubMed database, Elsevier database and Nature database were retrieved with key words “msenchymal stem cells (MSCs), microRNA (miRNA)” for papers published from December 2000 to November 2011 in Chinese and English. A total of 84 papers were retrieved, and 64 of which were included in the final analysis.
RESULTS AND CONCLUSION: MicroRNAs are endogenous 21-25nt short non-coding RNAs that can posttranscriptionally modulate gene expression by binding to the 3’-untranslated region of their target genes. Mesenchymal stem cells can be isolated from bone marrow, adipocyte tissue, cord blood and umbilical cord, and are capable of differentiating into multilineage cell types, such as osteoblasts, chondroblasts, adipocytes, myoblasts and neuroblasts. Recently, studies show that microRNAs play an important role in regulating proliferation, self-renewal and differentiation of mesenchymal stem cells by modulating expression of some specific transcription factors and genes related to the developmental process.

CLC Number:

R318

Chen Xue-bin, Xu Yin-sheng, Zhang Fang, Sheng Wang. MicroRNAs regulate self-renewal and differentiation of mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2012, 16(49): 9294-9300.

Figures/Tables 1

2.1 miRNA在间充质干细胞自我更新中的功能 miRNA可以调控干细胞自我更新，见表1。Xu等[19]的报道中，过表达miR-145会导致胚胎干细胞自我更新能力降低，miR-145可以通过抑制干细胞重组的重要因子sox2、oct4、klf-4的表达而降低胚胎干细胞干性。研究表明在鼠纤维细胞中过表达miR-302-367家族miRNA可以提高由Sox2、Klf4和Oct4诱导形成诱导全能干细胞的能力[20]，类似的研究表明miR-93和miR-106通过靶向调控转化生长因子β受体Ⅱ的表达[21]，miR-302/372通过抑制靶基因CDKN1A、RBL2、MECP2的表达提高了细胞向诱导全能干细胞的转化效率[22]，miR-34通过调控靶基因nanog、sox2、mycn的表达抑制了干细胞自我更新和重组[23]。 Giraud-Triboult等[24]比较了间充质干细胞和胚胎干细胞miRNA和mRNA的表达谱差异，发现在间充质干细胞中miR-148a和miR-20b表达下降，其靶基因EPAS1转录因子的表达增高，从而提高间充质干细胞形态特异性蛋白的表达，维持间充质干细胞的形态特征。Yu等[25]比较了年老猴子和年轻猴子骨髓间充质干细胞活性和miRNA表达的差异。与年轻猴子相比年老猴子骨髓间充质干细胞中端粒酶的表达下降了约65%，miRNA芯片分析结果显示在年老猴子的间充质干细胞中，miR-766和miR-558表达升高，miR-125b、let-7、miR-222、miR-199-3p、miR-23a和miR-221表达下降。类似的研究中Hackl等[26]发现miR-17、miR-19b、miR-20a和miR-106a在细胞老化模型中表达降低，这些miRNA对于间充质干细胞的自我更新具有重要作用。Riggi等[27]的研究发现EWS-FLI-1在胚胎干细胞中可以激活Oct4、Nanog、Sox2的表达，在间充质干细胞中过表达EWS-FLI-1能够提高Sox2的表达，同时降低miR-145的表达，而miR-145的一个靶基因就是Sox2，EWS-FLI-1和miR-145通过形成抑制性的反馈调节调控Sox2表达来调节干细胞的干性。Nie等[28]分析了miRNA与间充质干细胞凋亡之间的关系，作者发现在间充质干细胞中过表达miR-21、miR-23a和miR-210可以增强干细胞在低氧环境中的生存率，降低这些miRNA的表达会加快细胞凋亡。总之miRNA在间充质干细胞的自我更新过程中发挥着重要作用，随着人们对间充质干细胞自我更新认识的加深，miRNA在其过程中的作用也会越来越重要。 2.2 miRNA在间充质干细胞分化中的功能间充质干细胞在适当的条件下可以诱导分化为骨细胞、软骨细胞、肌肉细胞、神经细胞和脂肪细胞等，在间充质干细胞分化过程中伴有miRNA表达的变化，特异性miRNA在间充质干细胞分化过程中发挥着重要的调控作用。 2.2.1 miRNA对间充质干细胞成骨分化过程的调控间充质干细胞在向骨分化的过程中会受到血小板衍生生长因子、wnt、骨形态发生蛋白等多种信号途径的调节。Goff等[29]研究发现血小板衍生生长因子信号途径可以通过控制miRNA的表达来促使间充质干细胞向骨方向分化。Wnt途径的激活可以促进间充质干细胞的成骨分化，研究表明，miR-27通过调控靶基因APC的表达激活wnt信号途径[30]，miR-29通过抑制wnt信号途径的反向调控因子Dkk1、Kremen2、sFRP2激活wnt信号途径[31]，从而促使细胞向骨细胞方向分化。Runx2是成骨过程中一个重要的细胞因子，它可以激活与成骨相关基因的表达，其表达受到骨形态发生蛋白信号途径的调节，间充质干细胞中过表达miR-204和miR-335会抑制RUNX2的表达[13，32]，从而降低间充质干细胞向骨方向分化的能力。Mizuno等[33]研究表明miR-125b可以调控骨形态发生蛋白诱导的成骨过程，当miR-125的表达被抑制时，由骨形态发生蛋白诱导的碱性磷酸酶表达增多，会促进细胞成骨分化。Zhang等[34]研究发现miR-20a的靶基因是骨形态发生蛋白信号途径的反向调控因子PPARγ、Bambi和CRim1，在成骨过程中miR-20a表达增加，miR-20a通过调控骨形态发生蛋白-RUNX2信号途径而促使细胞向骨方向分化。 Gao等[35]通过生物芯片技术分析了骨髓间充质干细胞成骨诱导前后miRNA表达谱的变化，得到4个表达下降的miRNA(miR-31、miR-106a、miR-148a和miR-424)和3个表达升高的miRNA(miR-30c、miR-15b和miR-130b)，通过靶基因预测作者认为4个显著下降的miRNA靶基因可能是Runx2、CBFB、骨形态发生蛋白，而表达升高的miRNA靶基因可能是间充质干细胞特异性标记蛋白CD44、ITGB1、FLT1和间充质干细胞干性维持因子成纤维细胞生长因子2和CXCL12，这些miRNA共同作用促使了细胞向骨方向分化。类似研究中Schoolmeesters等[36]分析了间充质干细胞向骨分化前后miRNA表达谱的变化，发现miR-27a，miR-489在分化以后表达下调，miR-148b在成骨分化后表达下调，实验表明通过改变这3个miRNA在间充质干细胞中的表达，可以促使细胞在不加诱导剂的情况下向成骨方向分化。Eskildsen等[37]的研究表明在体外成骨过程中抑制miR-138的表达可以促进成骨特异性基因碱性磷酸酶的表达，促进间充质干细胞成骨分化，体内研究证实抑制miR-138的表达可以使成骨能力提高60%。在Kim等[38]的研究中，miR-196通过降低靶基因HOXC8的表达促使间充质干细胞向骨方向分化，在间充质干细胞中过表达miR-196a可以增强干细胞向骨分化的能力。总之，miRNA在间充质干细胞的成骨分化过程中发挥着重要的调控作用。 2.2.2 miRNA对间充质干细胞成软骨分化中的调控间充质干细胞向软骨分化过程也受到了miRNA的调控，在软骨分化过程中，早期细胞聚集是软骨细胞形成的一个必须步骤，Song等[39]发现miR-488在细胞聚集的早期高表达，在软骨分化以后表达降低，研究发现miR-488通过调节靶基因基质金属蛋白酶3的表达水平调控了软骨分化早期细胞聚集过程，促进了软骨分化的发生。 Miyaki等[40]分析了人骨髓来源的间充质干细胞和正常软骨细胞miRNA表达谱差异，结果显示miR-140在两种细胞中表达差异最显著。研究表明miR-140与正常软骨细胞基因表达有密切联系，在骨关节炎软骨或白细胞介素6诱导的条件下，软骨细胞中miR-140表达下降，miR-140表达降低会造成正常软骨细胞特异性蛋白表达失调。Yan等[6]利用生物芯片技术比较了大鼠骨髓间充质干细胞成软骨诱导后和正常软骨细胞的miRNA表达谱，在44个表达差异显著的miRNA中，检测了miR-29a和miR-29b在成软骨诱导过程中的调控作用，这两个miRNA的靶基因都是软骨特异性的蛋白collagenⅡ，过表达miR-29a和miR-29b会导致间充质干细胞软骨分化能力降低。在细胞成软骨诱导过程中，转化生长因子β信号通路起着非常重要的作用，Lin等[41]研究发现，miR-199a*通过抑制骨形态发生蛋白Ⅱ信号途径smad1蛋白的表达抑制了其下游sox9，collagenⅡ，COMP的表达，过表达miR-199a*减弱了间充质干细胞向软骨细胞分化的能力。Yang等[42]分析了鼠骨髓间充质干细胞软骨分化前后miRNA的表达谱变化。研究发现，过表达miR-145可以降低转化生长因子β信号途径下游与软骨分化相关的重要转录因子sox9的表达，抑制了间充质干细胞向软骨细胞的分化[14]。本实验室利用芯片检测人脐带间充质干细胞成软骨分化前后miRNA的表达情况，研究发现miR-485和miR-889可以通过调节sox9的表达来调控间充质干细胞成软骨分化的能力(数据未发表)。 2.2.3 miRNA对间充质干细胞成脂肪分化的调控近些年来，肥胖已成为困扰人类健康的一大问题，与脂肪相关的研究越来越多，miRNA对脂肪形成过程的调控也引起了人们的注意。Bengestrate等[43]分析人脂肪来源间充质干细胞成脂肪分化前后miRNA表达谱的变化，得到了4个表达明显调高的miRNA(miR-26a、miR-99a、miR-143和miR-455)以及3个表达明显下降的miRNA(miR-7、miR-130b、miR-92)，研究发现这些miRNA的表达与脂肪特性基因的表达密切相关。Nakanishi等[44]利用生物芯片和生物信息学手段分析了肥胖症大鼠肝脏脂肪细胞的miRNA表达谱，研究表明当细胞向脂肪细胞分化时miR-335表达增加，miR-335的表达与肪细胞特异性蛋白PPARγ、Ap2、FAS的表达密切相关，作者认为肝脏中miR-335表达水平可以作为检测肥胖症的一个标记。类似的研究中Ortega等[45]分析了成熟脂肪细胞和早期脂肪母细胞miRNA表达水平的差异，得到了50个表达差异明显的miRNA，这些在脂肪分化前后表达差异显著的miRNA可能会成为肥胖症或与肥胖症相关一些疾病的特异性标记物或治疗靶点。转化生长因子β信号途径可以抑制间充质干细胞向脂肪分化，Kim等[46]研究发现在间充质干细胞中过表达miR-21会抵消转化生长因子β引起成脂分化抑制的现象，研究表明miR-21的一个靶基因是转化生长因子β信号通路的受体转化生长因子β受体Ⅱ，而且miR-21的表达还会调控到smad3的磷酸化水平，miR-21通过转化生长因子β信号通路的抑制促进了间充质干细胞向脂肪分化。在Sun等[47]的研究中，let-7可以通过部分抑制细胞克隆形成和增殖相关转录因子HMGA2的表达来促进3T3L1细胞成脂肪分化，miR-143通过抑制Erk5蛋白的表达促进了细胞成脂肪分化[48]。Kim等[49]报道miR-27a在成脂诱导中表达降低，过表达miR-27a可以通过抑制PPARγ的表达调控细胞成脂分化能力。Wang等[50]的研究表明，在3T3L1细胞中稳定转染miR-17-92家族miRNA可以增加细胞成脂肪分化过程中三酰甘油的积累，实验证实miR-17-92通过靶向抑制Rb2/p130的表达增强了细胞成脂分化能力。研究表明miR-369-5p、miR-371、miR-138、miRNA-378/378*和miR-210等在干细胞成脂肪分化过程中都发挥着重要的调控作用[51-54]。 2.2.4 miRNA对间充质干细胞成肌肉分化的调控间充质干细胞在向肌肉分化的过程也受到miRNA的调控。Danielson等[55]研究表明miRNA的表达是间充质干细胞向肌肉细胞分化所必需的，实验比较了正常骨髓间充质干细胞和缺失Drosha和Dicer的间充质干细胞成肌肉分化的能力，发现突变的间充质干细胞不能分化为肌肉细胞。在Shan等[56]的研究中，作者检测了与肌肉细胞不接触共培养的间充质干细胞和用5-氮胞苷成肌肉诱导间充质干细胞中miRNA的表达变化，作者发现此过程中在肌肉细胞特异性蛋白开始表达的同时miR-143，miR-181，miR-206，miR-208表达增高，作者认为这些miRNA与肌肉分化密切相关。在Chen等[57]的研究中，miR-1和miR-133可以分别通过抑制靶基因HDAC4和SRF的表达来调控胚胎发育中骨骼肌相关基因的表达。Naguibneva等[58]的研究表明miR-181在肌肉细胞中高表达，实验证实miR-181的高表达可以抑制肌肉细胞分化的一个抑制因子Hox-A11的表达，从而促进细胞向肌肉细胞分化。 2.2.5 miRNA对间充质干细胞成神经分化的调控在间充质干细胞成神经分化过程中Notch信号途径起反向调节作用，Jing等[59]的研究发现，miR-9在神经诱导过程中表达增高，实验表明miR-9的靶基因是notch1蛋白，miR-9通过调节Notch信号途径增强了间充质干细胞向神经分化的能力。研究表明miR-206和miR-130a可以通过对靶基因TacI的调节来调控神经细胞中SP的合成与释放[60]。Maisel等[15]分析了间充质干细胞向神经分化前后mRNA和miRNA表达谱变化，研究显示HIF-1和miR-124a通过调节与氧相关的信号通路促进间充质干细胞向神经细胞的分化。Crobu等[61]分析了骨髓间充质干细胞成神经分化前后的miRNA表达谱差异，发现miR-31和miR-138在间充质干细胞中表达较高，而miR-21、miR-143、miR-23b在成神经诱导后表达升高。类似的研究中，miR-373、miR-584、miR-149、miR-91、miR-30c等在成神经诱导后的表达增高，这些神经诱导前后发生变化的miRNA与神经细胞分化有着密切的关系[62]。综上所述，miRNA在间充质干细胞向骨、软骨、脂肪、肌肉、神经等分化过程中起到重要的调控作用。 2.3 miRNA在间充质干细胞中的其他调控作用最近研究发现细胞来源的微泡(microvesicle)是细胞与细胞之间信息传导的一种新机制，这些微泡通过与靶细胞的融合来传递其中含有的基因信息。研究表明胚胎干细胞分泌的微泡中所含mRNA对造血干细胞的重编有调控作用[63]。在人体中，间充质干细胞可以通过释放一些细胞因子、激酶和细胞基质蛋白分子而修复受损组织。Collino等[64]从间充质干细胞分泌的微泡中检测到了多种miRNA，通过芯片分析发现有些miRNA在微泡中富集，间充质干细胞对其他组织细胞的调节和修复，有一部分是通过微泡中的miRNA发挥作用的。"

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