摘要
目的 探讨NUS1P1调控胶质瘤细胞增殖、迁移和侵袭的作用及机制。方法 运用小干扰RNA(siRNA)敲低胶质瘤细胞系U87和U251细胞中NUS1P1表达(分别设置NC组和si-NUS1P1组),应用细胞计数试剂盒8(CCK8)、细胞克隆形成实验、流式细胞术、划痕实验和Transwell实验检测细胞增殖、凋亡、迁移和侵袭能力;蛋白质印迹法检测凋亡相关蛋白、Wnt/β-catenin及AKT/mTOR信号通路中关键蛋白表达水平。组间比较采用独立样本t检验。结果在U87和U251细胞中,敲低NUS1P1 72h后,si-NUS1P1组细胞存活率均低于NC组,t值分别为3.39和4.33,P值分别为0.028和0.012。克隆形成实验结果显示,在U87细胞中,NC组和si-NUS1P1组细胞克隆数分别为(152.0±6.0)和(72.0±3.0)个,t=21.48,P<0.001;在U251细胞中,NC组和si-NUS1P1组细胞克隆数分别为(186.0±8.0)和(115.0±7.0)个,t=11.81,P<0.001。流式细胞术检测结果显示,U87和U251细胞敲低NUS1P1后细胞凋亡率均升高,si-NUS1P1组分别为(14.71±0.77)%和(37.55±1.09)%,NC组分别为(7.56±0.32)%和(27.00±0.71)%,差异均有统计学意义,均P<0.001。划痕愈合实验及Transwell迁移侵袭实验结果显示,敲低NUS1P1可抑制胶质瘤细胞迁移和侵袭能力,均P<0.001。蛋白质印迹法结果显示,在U87和U251细胞中,与NC组相比,si-NUS1P1组Bax、Caspase 9、Cleaved-Caspase 3和E-cadherin蛋白水平均升高,Bcl2、β-catenin、Wnt3a、N-cadherin、AKT/p-AKT和p-mTOR蛋白水平均降低,均P<0.05。结论 敲低NUS1P1可激活AKT/mTOR和Wnt/β-catenin信号通路,抑制胶质瘤细胞增殖、迁移和侵袭,诱导细胞凋亡,引起上皮-间质转化相关蛋白水平变化。
Objective To investigate the role and mechanism of NUS1P1in regulating the proliferation,migration,and invasion of glioma cells.Methods Small interfering RNA(siRNA)was used to knock down NUS1P1expression in glioma cell lines U87and U251(designated as the NC group and si-NUS1P1group,respectively).Cell Counting Kit-8(CCK-8),colony formation assay,flow cytometry,wound healing assay,and Transwell assay were employed to assess cell proliferation,apoptosis,migration,and invasion.Western blotting was used to detect the expression levels of apoptosis-related proteins and key proteins in the Wnt/β-catenin and AKT/mTOR signaling pathways.Comparisons between groups were performed using the independent samples t-test.Results At 72hours after NUS1P1knockdown in U87and U251 cells,the cell survival rate in the si-NUS1P1group was significantly lower than that in the NC group(t values were 3.39 and 4.33;Pvalues were 0.028and 0.012).The colony formation assay showed that the number of cell clones in the si-NUS1P1group was significantly reduced compared to the NC group[U87:(72.0±3.0)vs(152.0±6.0),t=21.48,P<0.001;U251:(115.0±7.0)vs(186.0±8.0),t=11.81,P<0.001].Flow cytometry results indicated a significant increase in the apoptosis rate after NUS1P1knockdown[U87si-NUS1P1:(14.71±0.77)%vs NC:(7.56±0.32)%;U251si-NUS1P1:(37.55±1.09)%vs NC:(27.00±0.71)%;both P<0.001].Furthermore,the wound healing and Transwell assays demonstrated that NUS1P1knockdown inhibited the migration and invasion abilities of glioma cells(both P<0.001).Western blot analysis revealed that,compared with the NC group,the si-NUS1P1group exhibited increased protein levels of Bax,Caspase9,Cleaved-Caspase3,and E-cadherin,and decreased levels of Bcl2,Wnt3a,β-catenin,N-cadherin,AKT/p-AKT,and p-mTOR in both U87and U251cells(all P<0.05).Conclusion Knockdown of NUS1P1can inhibit proliferation,migration,and invasion,while promoting apoptosis in glioma cells.These effects are associated with the modulation of epithelial-mesenchymal transition(EMT)-related proteins and the regulation of the AKT/mTOR and Wnt/β-catenin signaling pathways.
作者
李加美
姚志刚
杨洪安
马继伟
冯吉贞
LI Jiamei;YAO Zhigang;YANG Hongan;MA Jiwei;FENG Jizhen(Shandong Provincial Hospital Affiliated to Shandong First Medical University,Jinan,Shandong 250021,China)
出处
《中华肿瘤防治杂志》
北大核心
2025年第23期1418-1426,共9页
Chinese Journal of Cancer Prevention and Treatment
基金
山东省自然科学基金(ZR2023MH220)
