MicroRNA-26a的表达对急性髓系白血病细胞增殖的影响

Effect of microRNA-26a expression on proliferation of acute myeloid leukemia cells

目的观察构建has-miR-26a重组慢病毒的表达载体对急性髓系白血病细胞增殖能力的影响。方法构建人MicroRNA-26a慢病毒表达载体(LV-miR-26a-GFP)并感染人急性髓系白血病细胞系NB4;分为未感染组、感染组(感染LV-miR-26a-GFP)以及阴性对照组(感染LV-GFP);用real-time RT-PCR检测miR-26a在NB4细胞内的表达水平,Western blot检测PTEN的表达;用CCK-8法绘制细胞的生长曲线和软琼脂克隆形成实验检测细胞的增殖。结果成功构建miR-26a重组慢病毒的表达载体,感染白血病细胞NB4后,能够成功过表达miR-26a。生长曲线的结果显示与感染LV-GFP阴性对照组和未感染组相比,LV-miR-26a-GFP感染组细胞的增殖速度显著增加(P<0.01),软琼脂克隆形成实验证实了感染LV-miR-26a-GFP的NB4细胞的克隆形成能力(GFP+克隆数为75+10)与阴性对照组(感染LV-GFP)相比(GFP+克隆数为26+5)显著增强(P<0.05)。在NB4细胞中过表达miR-26a后,其靶基因PTEN的表达明显被抑制。结论 MicroRNA-26a的表达可增强急性髓系白血病细胞的增殖能力,促进白血病细胞的增殖。

Objective To construct the recombinant lentivirus vector expressing has-miR-26a and to observe the effect of overexpression of miR-26a on acute myeloid leukemia cell proliferation.Methods The lentiviral vector of MicroRNA-26a（LV-miR-26a-GFP） was constructed and transfected into human myeloid leukemia cells.The cells were divided into non-transfected group,transfected group（transfected with LV-miR-26a-GFP） and negative control group（transfected with LV-GFP）.The expression level of miR-26a was measured by RT-PCR.The expression of targeted gene PTEN was tested by Western blot.The cell growth curves and the effects of the miR-26a on the ability of cell proliferation were respectively analyzed by CCK-8 and soft agar colony formation assay.Results The expression vectors of miR-26a lentivirus have been successfully achieved.After transfected acute myeloid leukemia cell NB4,we successfully achieved the over expression of miR-26a.Compared with non-transfected group and negative control group,the proliferation rate of cells transfected with LV-miR-26a-GFP group was significantly increased（P0.01）.Soft agar colony formation assay suggested that colony formation ability of NB4 cells transfected with LV-miR-26a-GFP group（the number of GFP＋ clones： 75＋10） was significantly enhanced,compared with negative control group（the number of GFP＋ clones： 26＋5）（P0.05）.The miR-26a overexpression can inhibit the expression of target gene PTEN in NB4 cells.Conclusions MicroRNA-26a inhibits the expression of target gene PTEN,enhances proliferation of acute leukemia cell.