In Vitro Functional Study of miR-126 in Leukemia
In Vitro Functional Study of miR-126 in Leukemia
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DOI：
10.1007/978-1-60761-863-8_13
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Chen, Jianjun
中科院分区：
文献类型：
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作者：
Li, Zejuan;Chen, Jianjun
微小RNA（miRNAs，miRs）被认为是多种癌症（包括白血病）的重要调节因子。在对52例急性髓性白血病（AML）样本中的435种人类miRNA进行的大规模miRNA表达谱分析中，我们发现miR-126及其在生物发生中的次要对应物，即miR-126*，在核心结合因子（CBF）AML（包括t（8;21）/AML 1-ETO和inv（16）/CBFB-MYH 11样本）中特异性异常过表达。我们的体外功能获得和丧失实验表明，miR-126的强制表达抑制了AML细胞的凋亡并增加了AML细胞的活力，而当miR-126的内源性表达被敲低时，观察到相反的效果。此外，通过体外集落形成/再铺板测定，我们证明了miR-126的强制表达单独增强小鼠正常骨髓祖细胞的增殖和集落形成/再铺板能力，特别是与AML 1-ETO（t（8;21）产生的融合基因）协同增强。因此，我们的数据表明，miR-126可能在CBF白血病的发展中发挥关键作用。在本章中，描述了用于研究白血病中miR-126的材料和方案。
MicroRNAs (miRNAs, miRs) are postulated to be important regulators in various cancers, including leukemia. In a large-scale miRNA expression profiling analysis of 435 human miRNAs in 52 acute myeloid leukemia (AML) samples, we found that miR-126 and its minor counterpart in biogenesis, namely, miR-126*, were specifically aberrantly overexpressed in core binding factor (CBF) AMLs including both t(8;21)/AML1-ETO and inv(16)/CBFB-MYH11 samples. Our in vitro gain- and loss-of-function experiments showed that forced expression of miR-126 inhibited apoptosis and increased the viability of AML cells, whereas the opposite effect was observed when endogenous expression of miR-126 was knocked down. In addition, through in vitro colony-forming/replating assays, we demonstrated that forced expression of miR-126 enhanced proliferation and colony-forming/replating capacity of mouse normal bone marrow progenitor cells alone and particularly, in cooperation with AML1-ETO, a fusion gene resulting from t(8;21). Thus, our data shows that miR-126 may play a critical role in the development of CBF leukemias. In the present chapter, the materials and protocols for the study of miR-126 in leukemia are described.