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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: To investigate the effect of microRNA-199a-3p (miR-199a-3p) on human osteosarcoma cell apoptosis. METHODS: Human osteosarcoma cells (U2-OS) were transfected with the synthetic miR-199a-3p mimics and negative cells were transfected with negative control sequences (NC mimics). The expression of miR-199a-3p in each group was detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) after transfection. Western blot was used to detect the expression of myeloid leukemia-1 (MCL-1) And the cleavage of poly (ADP-ribose) polymerase (PARP). The apoptosis rate of each group was detected by flow cytometry, and the experimental results were statistically analyzed. Results: Compared with the control group, miR-199a-3p mimics transfected cells showed significantly increased expression of miR-199a-3p, reduced expression of MCL-1 protein and increased cleavage of PARP protein, Cell apoptosis rate increased, the difference was statistically significant (P <0.05). Conclusion: miR-199a-3p can effectively promote the apoptosis of osteosarcoma cells.