microRNA-515-5p调控c-MYC-p53信号轴对神经母细胞瘤细胞增殖、侵袭和迁移的影响

Effects of microRNA-515-5p on Proliferation, Invasion and Migration of Neuroblastoma Cells by Regulating c-MYC-p53 Signal Axis

目的:探讨microRNA-515-5p(miR-515-5p)对神经母细胞瘤(NB)SK-N-BE(2)和SK-N-AS细胞增殖、侵袭、迁移的影响及分子机制。方法:采用生物信息学技术结合多种软件预测方法,检测miR-515-5p在NB细胞中的表达。在NB细胞中瞬时转染miR-515-5p mimics,应用CCK8法和克隆形成实验、Transwell侵袭小室实验和体外划痕实验检测miR-515-5p对NB细胞转染后的增殖、侵袭和迁移的影响。采用Western免疫印迹(Western Blot)检测miR-515-5p对c-MYC-p53信号轴的调控机制。结果:与对照组比较,转染miR-515-5p mimics的细胞集落平均形成率降低,细胞活性下降,且细胞侵袭和迁移能力降低(P<0.05)。Western Blot检测结果显示miR-515-5p抑制c-MYC蛋白表达,但可促进P-p53活化(P<0.05),对p53的表达无影响。结论:miR-515-5p通过调控c-MYC-p53信号轴抑制NB细胞的增殖、侵袭和迁移能力,可能作为NB分子治疗的潜在靶点。

Objective: To probe into the effects of microRNA-515-5 p(miR-515-5 p) on proliferation, invasion, migration and molecular mechanisms of SK-N-BE(2) and SK-N-AS of neuroblastoma(NB) cells. Methods: Bioinformatics techniques combined with multiple software prediction methods were used to detect the expression of miR-515-5 p in NB. The miR-515-5 p mimics were transiently transfected in NB cells, and effects of miR-515-5 p on the proliferation, invasion and migration of NB cells after transfection were examined by CCK8 method and colony-forming unit assay, Transwell invasion cell assay and in vitro scratch assay. Western Blot was used to detect the regulatory mechanism of miR-515-5 p on the c-MYC-p53 signal axis. Results: Compared with the control group, cells transfected with miR-515-5 p mimics had the reduced mean colony formation rate, decreased cell activity, and reduced cell invasion and migration ability(P<0.05). Western Blot showed that miR-515-5 p inhibited the expression of c-MYC protein, yet promoted the activation of P-p53(P<0.05), and had no effect on the expression of p53. Conclusion: The miR-515-5 p inhibits the proliferation, invasion and migration of NB cells by regulating the c-MYC-p53 signaling axis, which may be the potential target for molecular therapy of NB.