MicroRNA-199a-3p对人骨肉瘤细胞凋亡的影响

Effects of microRNA-199a-3p on apoptosis of human osteosarcoma cells

目的:探讨microRNA-199a-3p(miR-199a-3p)对人骨肉瘤细胞凋亡的影响。方法:用合成的成熟miR-199a-3p序列模拟物(miR-199a-3p mimics)转染人骨肉瘤细胞(U2-OS),以阴性对照物序列(NC mimics)转染细胞作为阴性对照。转染后应用实时定量逆转录聚合酶链反应(qRT-PCR)检测各组细胞miR-199a-3p的表达量,Western blot法检测各组细胞中髓样细胞白血病-1(MCL-1)蛋白的表达水平及多聚腺苷二磷酸核糖聚合酶(PARP)的剪切情况,利用流式细胞仪检测各组细胞的凋亡率,并对实验结果进行统计学分析。结果:与对照组相比,转染miR-199a-3p mimics的实验组细胞中,miR-199a-3p的表达量明显升高,MCL-1蛋白的表达降低,PARP蛋白的剪切水平增加,细胞的凋亡率增高,差异均有统计学意义(P<0.05)。结论:miR-199a-3p可以有效促进骨肉瘤细胞的凋亡。

Objective：MicroRNA-199a-3p（miR-199a-3p） is down-regulated in many kinds of tumors,including osteosarcoma. This study was aimed to investigate the effect of miR-199a-3p on apoptosis of human osteosarcoma cells（U2-OS）. Methods：Human osteosarcoma cells（U2-OS） were transfected with miR-199a-3p mimics or negative control mimics（NC mimics） as negative control. MiR-199a-3p expression level was detected by quantitative real-time reverse transcriptase polymerase chain reaction（qRT-PCR）. Expression level of myeloid cell leukemia-1（MCL-1） protein and cleavage of poly ADP-ribose polymerase（PARP） were detected by Western blot. Cell apoptosis was detected by flow cytometry. Results：Compared with control group,miR-199a-3p expression level of experiment group was significantly up-regulated,MCL-1 protein expression was reduced,cleavage level of PARP was increased and cell apoptosis rate of experiment group was significantly increased. The differences between experimental and control groups have statistical significance（P 0.05）. Conclusion：MiR-199a-3p can dramatically promote apoptosis of human osteosarcoma cells.