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miRNA-93在急性淋巴细胞白血病中的表达

         

摘要

AIM:To investigate the expression of microRNA (miRNA)-93 in acute lymphocytic leukemia (ALL) and its effect on the proliferation of acute T-cell leukemia Jurkat cells.METHODS:The expression of miRNA-93 in the bone marrow samples of patients with ALL was measured by real-time PCR.After down-regulation of miRNA-93 by transfection with miRNA-93 inhibitor in the Jurkat cells,the cell viability,cell proliferation and cell cycle distribution were detected by CCK-8 assay,EdU assay and flow cytometry,respectively.Furthermore,the protein levels of cell cycle-related molecules such as cyclin D1,cyclin-dependent kinase 4 (CDK4),phosphorylation retinoblastoma (Rb) and P27 were measured by Western blot.RESULTS:miRNA-93 was highly expressed in the patients with ALL,and the expression level was highest in the high risk patients.Down-regulation of miRNA-93 inhibited Jurkat cell viability,arrested cell cycle in G1/S transition.In addition,the protein levels of cyclin D1,CDK4 and p-Rb were significantly decreased,the protein expression of P27 was increased in Jurkat cells trasfected with miRNA-93 inhibitor.CONCLUSION:miRNA-93 expression is increased in ALL patients.Down-regulation of miRNA-93 restrains cell proliferation in the acute T cell leukemia cell line Jurkat via regulating cell cycle-related molecules.%目的:探讨微小RNA(miRNA)-93在急性淋巴细胞白血病中的表达情况及其对急性T细胞白血病细胞株Jurkat增殖的影响和潜在作用机制.方法:Real-time PCR技术检测急性白血病患者骨髓样本中miRNA-93的表达水平.转染miRNA-93 inhibitor下调Jurkat细胞中miRNA-93的表达,分别采用CCK-8法、EdU法和流式细胞术检测细胞活力、增殖及周期,Western blot检测周期相关调控因子cyclin D1、细胞周期蛋白依赖性激酶4(CDK4)、p-Rb及P27的蛋白表达水平.结果:miRNA-93在急性白血病患者中呈现高表达,且在高危患者中表达水平最高;沉默miRNA-93后,Jurkat细胞的活力下降并出现明显的G1/S期阻滞,同时细胞中cyclin D1、CDK4、p-Rb的蛋白水平显著降低,而P27的蛋白水平显著升高.结论:miRNA-93在急性淋巴细胞白血病中表达显著升高;沉默miRNA-93可通过调控周期相关因子的表达抑制急性T细胞白血病细胞株Jurkat的增殖.

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