Analysis of miRNA expression patterns in tetracycline (Tet) inducible Scl/tTA-BCR/ABL (Scl-BA) transgenic mouse hematopoietic stem-progenitor cells.

To further define the roles of miRNAs in the pathogenesis of CML, we have employed miRNA microarray expression profiling as a discovery platform to identify miRNAs with the potential to drive or accelerate CML progression. Hematopoietic stem-progenitor cells miRNA profiles of BA mice after BCR/ABL expression induced for 0 week and 3 weeks were generated by miRNA microarray. Expression of six miRNAs (miR-126a-3p, miR-130a-3p,miR-142a-5p, miR-142a-3p, miR-29c-3p and miR-30c-5p) from this signature was quantified in the same RNA samples by real-time PCR, confirming low variability in BCR/ABL expressing cells. Hematopoietic stem-progenitor cells (lin-c-kit+sca1+, LSK) of BA mice after BCR/ABL expression induced for 0 week and 3 weeks were sorted by FACS, LSK cells from three mice were mixed in each group.

为进一步阐明微小RNA（miRNAs）在慢性髓系白血病（chronic myeloid leukemia, CML）发病机制中的作用，本研究采用miRNA微阵列表达谱作为发现平台，以识别具有驱动或加速CML进展潜力的miRNAs。本研究通过miRNA微阵列，获取了BCR/ABL融合基因诱导表达0周及3周的BA小鼠造血干祖细胞的miRNA表达谱。随后，本研究通过实时定量PCR对该特征表达谱中的6种miRNAs（miR-126a-3p、miR-130a-3p、miR-142a-5p、miR-142a-3p、miR-29c-3p及miR-30c-5p）在相同RNA样本中的表达水平进行了定量验证，结果证实BCR/ABL阳性细胞中的表达变异度较低。本研究通过荧光激活细胞分选术（FACS）分选出BCR/ABL诱导表达0周及3周的BA小鼠造血干祖细胞（lin⁻c-kit+sca1+, LSK），并将每组3只小鼠的LSK细胞混合后用于后续实验。