miR-125b在儿童AML中的表达及其反义寡核苷酸对白血病细胞的作用

Expression of miR-125b in pediatric acute myeloid leukemia and the effect of anti-miR-125b oligonucleotide on proliferation of human leukemic cells

目的:研究儿童急性髓细胞白血病(AML)患者骨髓细胞中miR-125b的表达及miR-125b为靶标的反义寡核苷酸对人白血病细胞的作用。方法:采用基因芯片和实时荧光定量PCR技术(qRT-PCR)检测儿童AML治疗前后骨髓细胞中miR-125b的表达。采用电转法将与miR-125b序列互补的反义寡核苷酸转染HL-60细胞,细胞计数试剂盒(CCK-8)检测转染后24 h、48 h、72 h、96 h细胞增殖情况。结果:芯片结果显示miR-125b在儿童AML中表达明显增高,约为正常的12倍,qRT-PCR结果进一步证实了miR-125b在儿童AML中异常高表达,同时还发现miR-125b在儿童AML部分缓解的患者骨髓细胞中表达下降,在完全缓解患者骨髓细胞中降至正常水平。CCK-8结果显示,针对miR-125b的反义寡核苷酸能有效抑制白血病细胞的增殖,与对照组相比有显著差异(P<0.01)。结论:miR-125b在儿童AML中可能起"癌基因"作用,以miR-125b为靶标的反义寡核苷酸可能为儿童AML治疗提供新的方法。

AIM： To investigate the expression of miR-125b in pediatric acute myeloid leukemia（AML）,and to explore the inhibitory effect of anti-miR-125b oligonucleotide on human leukemic cells. METHODS： The expression of miRNAs in pediatric AML bone marrow cells was analyzed by gene microarray and real-time quantitative PCR.HL-60 cells were transfected with anti-miR-125b,which was complementary to miR-125b in sequence,and the viability of HL-60 cells was measured by CCK-8 assay 24 h,48 h,72 h and 96 h after electroporation. RESULTS： The expression levels of miR-125b in the cells of newly diagnosed pediatric AML patients was almost 12 times as high as that in the cells of idiopathic thrombocytopenic purpura cases.However,the decreased levels of miR-125b in the cells of partial remission patients and returned to normal in the cells of completely remission patients were observed.At the same time,the growth rate of the cells treated with anti-miR-125b oligonucleotide was obviously decreased compared with that of control cells. CONCLUSION： miR-125b may take effect as an oncogene in pediatric AML.Anti-miR-125b oligonucleotide may be useful as a new drug for treating pediatric AML.