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目的研究microRNA-21反义寡核苷酸对地西他滨(DCA)抗白血病效应的影响及可能的机制。方法将microRNA-21反义核苷酸(AMO)和无义寡核苷酸(SCR)通过脂质体转染导入U937细胞,实时荧光定量PCR(qRTPCR)验证转染效率,再分别与3种浓度的DCA(0.5μmol/L、2.0μmol/L、4.0μmol/L)作用48 h。采用Annexin V/PI法检测凋亡,通过流式细胞仪检测CD14和CD11b的平均荧光强度(MFI)和细胞周期。结果 AMO转染组的microRNA-21表达(0.67±0.09)低于空白组(1.10±0.06)和SCR转染组(1.04±0.08),差异有统计学意义(P<0.01)。AMO转染组的U937细胞DCA的IC50低于空白组和SCR转染组,差异均有统计学意义(P<0.01)。同一浓度下,AMO组的早期凋亡率、CD14 MFI、CD11b MFI、细胞Sub-G1期均高于同一浓度药物作用的空白组和SCR组,差异均有统计学意义(P<0.01)。结论 microRNA-21 AMO能显著促进DCA体外抗白血病效应,其机制可能与作用于细胞周期和分化抗原的改变有关。
Objective To study the effect of microRNA-21 antisense oligonucleotide on the anti-leukemia effect of decitabine (DCA) and its possible mechanism. Methods MicroRNA-21 antisense oligonucleotide (AMO) and non-sense oligonucleotide (SCR) were transfected into U937 cells by lipofectamine. The efficiency of transfection was verified by qRTPCR, Concentration of DCA (0.5μmol / L, 2.0μmol / L, 4.0μmol / L) for 48 h. Apoptosis was detected by Annexin V / PI assay. The mean fluorescence intensity (MFI) and cell cycle of CD14 and CD11b were detected by flow cytometry. Results The expression of microRNA-21 in AMO transfected group was significantly lower than that in blank transfected group (1.10 ± 0.06) and SCR transfected group (1.04 ± 0.08) (0.67 ± 0.09, P <0.01). The IC50 of DCA in U937 cells in AMO transfection group was lower than that in blank transfection group and SCR transfection group (P <0.01). At the same concentration, the early apoptotic rate, the CD14 MFI, the CD11b MFI and the cell sub-G1 phase in AMO group were all higher than those in blank group and SCR group with the same concentration of drug (P <0.01). Conclusion MicroRNA-21 AMO can significantly promote the anti-leukemia effect of DCA in vitro, which may be related to the changes of cell cycle and differentiated antigen.