MicroRNA-193 Pro-Proliferation Effects for Bone Mesenchymal Stem Cells After Low-Level Laser Irradiation Treatment Through Inhibitor of Growth Family, Member 5
MicroRNA-193 Pro-Proliferation Effects for Bone Mesenchymal Stem Cells After Low-Level Laser Irradiation Treatment Through Inhibitor of Growth Family, Member 5
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MicroRNA-193 通过生长抑制剂家族成员 5 进行低水平激光照射处理后对骨间充质干细胞的促增殖作用
DOI：
10.1089/scd.2011.0695
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发表时间：
2012-09-01
影响因子：
4
通讯作者：
Zhang, Hao
中科院分区：
文献类型：
--
作者：
Wang, Jue;Huang, Weicong;Zhang, Hao
骨髓间充质干细胞（mesenchymal stem cells，MSCs）的增殖能力增强有助于细胞治疗的临床转化。低强度激光照射（LLLI）已被证明是调节MSC增殖。microRNAs（miRNAs）参与干细胞的多种病理生理过程，但miRNAs在LLLI促进MSC增殖中的作用尚不清楚。我们发现LLLI处理后MSCs的增殖水平和细胞周期相关基因以时间依赖的方式增加。微阵列分析揭示了差异调节的miRNAs亚群，这些动态变化通过LLLI后的定量实时聚合酶链反应（qRT-PCR）证实。miR-193是上调最高的miRNA，其变化与细胞增殖水平相关。获得-丧失功能实验表明，miR-193可调控人和大鼠MSCs的增殖，但不影响其凋亡和分化水平。阻断miR-193可抑制LLLI诱导的MSC增殖。通过qRT-PCR，我们发现miR-193，特别是，调节细胞周期蛋白依赖性激酶2（CDK 2）的表达。生物信息学分析和荧光素酶报告基因分析表明，生长抑制因子家族成员5（inhibitor of growth family，member 5，ING 5）是miR-193调控细胞增殖和CDK 2活性的最佳靶点，其mRNA和蛋白水平均受miR-193调控。小干扰RNA（siRNA）抑制ING 5可上调MSCs的增殖和CDK 2的表达。总之，这些结果强烈表明，miR-193在响应LLLI刺激的MSC增殖中起关键作用，这可能适合于临床应用的治疗操作。
The enhanced proliferation of mesenchymal stem cells (MSCs) can be helpful for the clinical translation of cell therapy. Low-level laser irradiation (LLLI) has been demonstrated as regulating MSC proliferation. MicroRNAs (miRNAs) are involved in various pathophysiologic processes in stem cells, but the role of miRNAs in the LLLI-based promotion of MSC proliferation remains unclear. We found that the proliferation level and cell cycle-associated genes in MSCs were increased after LLLI treatment in a time-dependent manner. Microarray assays revealed subsets of miRNAs to be differentially regulated, and these dynamic changes were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) after LLLI. miR-193 was the most highly up-regulated miRNA, and the change in it was related with the proliferation level. Gain-loss function experiments demonstrated that miR-193 could regulate the proliferation of MSCs, including human's and rat's, but could not affect the apoptosis and differentiation level. Blockade of miR-193 repressed the MSC proliferation induced by LLLI. By qRT-PCR, we found that miR-193, in particular, regulated cyclin-dependent kinase 2 (CDK2) expression. Bioinformatic analyses and luciferase reporter assays revealed that inhibitor of growth family, member 5 (ING5) could be the best target of miR-193 to functionally regulate proliferation and CDK2 activity, and the mRNA and protein level of ING5 was regulated by miR-193. Furthermore, the ING5 inhibited by small interfering RNA (siRNA) could up-regulate the proliferation of MSCs and the expression of CDK2. Taken together, these results strongly suggest that miR-193 plays a critical part in MSC proliferation in response to LLLI stimulation, which is potentially amenable to therapeutic manipulation for clinical application.