miR-17-5p对神经母细胞瘤细胞增殖和迁移能力影响

EFFECT OF MIR-17-5P ON THE PROLIFERATION AND MIGRATION OF NEUROBLASTOMA CELLS

目的探讨miR-17-5p对神经母细胞瘤(NB)细胞增殖和迁移能力的影响及其相关机制。方法用miR-17-5p mimic和miR-17-5p inhibitor及二者阴性对照NC mimics和NC inhibitor分别转染SH-SY5Y细胞,应用实时荧光定量PCR(RT-PCR)方法检测转染后各组细胞内miR-17-5p的表达水平;利用CCK-8实验、细胞菌落形成实验、Transwell小室检测miR-17-5p对细胞增殖和迁移能力的影响;利用生物信息学网站预测miR-17-5p在10号染色体张力蛋白同源物磷酸脂酶(PTEN)中的结合位点;Western blot方法检测各组细胞中PTEN、蛋白激酶B(AKT)和磷酸化AKT(p-AKT)蛋白的表达情况。结果与NC mimics组相比,miR-17-5p mimics组SH-SY5Y细胞miR-17-5p表达、细胞菌落数量、细胞增殖及细胞迁移能力均升高,而PTEN蛋白表达降低、p-AKT/AKT蛋白表达比值升高;与NC inhibitor组相比,miR-17-5p inhibitor组SH-SY5Y细胞miR-17-5p表达、细胞菌落数量、细胞增殖及细胞迁移能力均降低,而PTEN蛋白表达升高、p-AKT/AKT蛋白表达比值降低,差异均有显著意义(t=2.940~12.990,F=23.07~408.47,P<0.05)。结论miR-17-5p具有促进NB细胞增殖和迁移的作用,其机制可能是通过下调PTEN表达激活PI3K/AKT信号通路实现的。

Objective To investigate the effect of miR-17-5p on the proliferation and migration of neuroblastoma(NB)cells and related mechanisms.Methods SH-SY5Y cells were transfected with miR-17-5p mimic,miR-17-5p inhibitor,and their negative controls NC mimics and NC inhibitor,respectively.Quantitative real-time PCR was used to measure the expression level of miR-17-5p in each group after transfection;CCK-8 assay,cell colony formation experiment,and Transwell chamber were used to observe the effect of miR-17-5p on cell proliferation and migration;a bioinformatics website was used to predict the binding site of miR-17-5p to phosphatase and tensin homolog(PTEN)on chromosome 10;Western blot was used to measure the protein expression of PTEN,protein kinase B(AKT),and phosphorylated AKT(p-AKT)in cells.Results Compared with the NC mi-mics group,the miR-17-5p mimics group had significant increases in the expression of miR-17-5p in SH-SY5Y cells,the number of cell colonies,and the proliferation and migration abilities of cells,a significant reduction in the protein expression of PTEN,and a significant increase in p-AKT/AKT ratio.Compared with the NC inhibitor group,the miR-17-5p inhibitor group had significant reductions in the expression of miR-17-5p in SH-SY5Y cells,the number of cell colonies,and the proliferation and migration abilities of cells,a significant increase in the protein expression level of PTEN,and a significant reduction in p-AKT/AKT ratio(t=2.940-12.990,F=23.07-408.47,P<0.05).Conclusion This study shows that miR-17-5p can promote the proliferation and migration of NB cells,possibly by downregulating the expression of PTEN to activate the PI3K/AKT signaling pathway.