NPM1促进骨肉瘤细胞迁移、增殖、侵袭的体外研究

NPM1 promotes the migration,proliferation,and invasion of osteosarcoma cells in vitro

目的 探讨核仁磷酸蛋白1 (NPM1)对骨肉瘤细胞(143B及U2细胞)恶性表型迁移、增殖和侵袭能力的影响。方法 采用Oncomine数据库查询NPM1在骨肉瘤中的表达水平;构建重组慢病毒载体,制成过表达NPM1慢病毒(Lv/ShNPM1)、阴性对照慢病毒(Lv/negative)和沉默NPM1慢病毒(Lv/ShNPM1)。分别转染人源骨肉瘤细胞系(143B及U2细胞),分为Lv/NPM1组、Lv/ShNPM1组、NC (Lv/negative)组。运用Western blot法检测各组别中NPM1蛋白的表达水平。分别运用CCK8法、Wound healing法、Transwell invasion法检测各组细胞的增殖能力、迁移能力和侵袭能力。结果 Oncomine数据库查询结果显示,NPM1在骨肉瘤中呈高表达水平;检测各组中NPM1蛋白表达的结果显示:与NC组比较,Lv/ShNPM1组中NPM1蛋白表达水平降低,差异具有统计学意义(P<0.05);U2-OS细胞中NC组、Lv/NPM1组和Lv/ShNPM1组细胞迁移率分别为(49.7±3.3)%、(64.1±7.4)%、(24.3±6.6)%,143B细胞分别为(63.5±4.3)%、(75±3.7)%、(30.1±5.9)%,两种细胞Lv/ShNPM1组迁移率明显低于NC组,差异均有统计学意义(P<0.05);U2-OS细胞中NC、Lv/NPM1组、Lv/ShNPM1组侵袭细胞数分别为98±26.3、164±41、38±14.6,143B细胞分别为104±19.1、191±32.4、41±27.6,两种细胞Lv/ShNPM1组侵袭细胞数明显少于NC组,差异均有统计学意义(P<0.05);CCK8实验结果显示143B和U2-OS细胞Lv/ShNPM1组增殖能力明显低于NC组,差异均有统计学意义(P<0.05)。结论 NPM1能够体外增强骨肉瘤细胞迁移、增殖和侵袭能力。

Objective To investigate the effect of nucleophosmin 1(NPM1)on the migration,proliferation,and invasion of osteosarcoma cells(143B and U2-OS cells).Methods Bioinformatic prediction was performed to investigate the expression level of NPM1 in osteosarcoma using Oncomine database.Recombinant lentivirus vector was constructed to prepare overexpressed NPM1 lentivirus(Lv/ShNPM1),negative control lentivirus(Lv/negative),and silent NPM1 lentivirus(Lv/ShNPM1),which were transfected into osteosarcoma cells(143B and U2-OS cells),respectively,as Lv/NPM1 group,Lv/ShNPM1 group,NC(Lv/negative)group.Western blot was used to detect the expression level of NPM1 protein in each group.CCK8 method,Wound healing method,and Transwell invasion method were used to detect the proliferation,migration,and invasion ability of cells in each group.Results Bioinformatic prediction showed that NPM1 protein was over-expressed in osteosarcoma cells.Western blot results revealed that the expression levels of NPM1 protein was significantly lower in Lv/ShNPM1 group than NC(Lv/negative)group.In U2-OS cells,the cell migration rates in NC(Lv/negative)group,Lv/NPM1 group,and Lv/ShNPM1 group were(49.7±3.3)%,(64.1±7.4)%,and(24.3±6.6)%,respectively;in 143B cells,the cell migration rates were(63.5±4.3)%,(75±3.7)%,and(30.1±5.9)%,respectively;the cell migration rates in Lv/ShNPM1 group were significantly lower than those in NC(Lv/negative)group(P<0.05).In U2-OS cells,the number of invasive cells in NC(Lv/negative),Lv/NPM1,and Lv/ShNPM1 groups were 98±26.3,164±41,and 38±14.6,respectively;in 143B cells,the number of invasive cells were 104±19.1,191±32.4,and 41±27.6,respectively;the number of invasive cells in Lv/ShNPM1 group were significantly lower than those in NC(Lv/negative)group(P<0.05).CCK8 experiment results showed that the proliferation ability of 143B and U2-OS cells in Lv/ShNPM1 group was significantly lower than that in NC(Lv/negative)group(P<0.05).Conclusion NPM1 promotes the migration,proliferation,and invasion of osteosarcoma cells in vitro.