miR-16促进Neuro2a细胞的凋亡

Overexrpession of miR-16 promotes apoptosis in Neuro2a cells

目的检测过表达miR-16对小鼠Neuro2a细胞周期和凋亡的影响,探讨miR-16在肿瘤发病过程中的可能机制。方法 构建能够在细胞中过表达miR-16的真核表达载体pcDNA3.1-miR-16,利用Real-time PCR验证了表达载体的过表达效应。应用pcDNA3.1-miR-16载体转染Neuro2a细胞,利用流式细胞术和TUNEL法检测Neuro2a细胞周期及凋亡改变情况。结果 转染过表达pcDNA3.1-miR-16载体后,成熟的miR-16表达与对照组相比明显升高(P<0.05)。在Neuro2a细胞中过表达miR-16后,用流式细胞术检测发现,Neuro2a细胞周期未见明显改变,与对照组相比,凋亡细胞数量有显著增加;进一步通过TUNEL法验证,发现miR-16的过表达能够促进Neuro2a的细胞凋亡。结论 过表达miR-16能够显著促进Neuro2a细胞凋亡而对细胞周期并无影响,提示miR-16可能对肿瘤治疗起作用。

Objective To investigate the role of miR-16 in cell cycle and apoptosis in Neuro2a cells and explore the possible function of miR-16 in tumor.Methods The overexpression plasmid pcDNA3.1-miR-16 of miR-16 was cloned and introduced into Neuro2a cells.Real-time PCR was chosen to test the overexpression of miR-16.Cell cytometry was used to study the role of miR-16 in cell cycle and apoptosis,and apoptosis cells were quantitatively examined by TUNEL.Results After tranfection of pcDNA3.1-miR-16,the mature miR-16 was significantly overexpressed（P0.05）.Cell cytometry results showed that the cell cycle didn＇t change after miR-16 overexpression while cell apoptosis were found and confirmed by TUNEL.Conclusion There was significant apoptosis of Neuro2a cell after overexpression of miR-16,the result suggested miR-16 may have potential significane in treatment of tumor.