敲减NEDD1表达抑制肺腺癌细胞增殖与迁移

目的:探讨神经前体细胞表达发育下调蛋白1(NEDD1)在肺癌发生和发展中的作用。方法:采用免疫组化实验检测NEDD1在肺癌组织中的表达情况,结合数据库资料分析NEDD1在肺癌组织与癌旁正常组织中的表达有无差异,并分析NEDD1的表达水平与肺癌预后的相关性;敲减NEDD1在A549细胞的表达后用平板集落形成实验检测集落形成能力的变化,流式细胞术检测细胞凋亡和细胞周期分布的情况,Transwell小室实验检测对细胞的迁移能力的影响,Western blot检测细胞周期相关蛋白的表达,并分析NEDD1与周期蛋白依赖性激酶2(CDK2)表达的相关性。结果:与正常肺组织相比,肺癌组织中NEDD1表达水平升高,Kaplan-Meier曲线预后分析显示NEDD1高表达者预后差,NEDD1低表达者预后较好;光镜下观察可见敲减NEDD1表达的A549细胞增殖速度明显减慢,平板集落形成能力降低(P<0.05);与对照组相比,敲减NEDD1表达促进A549细胞凋亡,抑制细胞的迁移;与对照组相比,敲减NEDD1表达后G 1期细胞的比例增加,细胞阻滞于G 1/S期,Western blot检测周期相关蛋白水平可见p-Rb和cyclinD1蛋白表达水平降低,p-Chk1、p-Chk2和p-p53蛋白表达水平升高(P<0.05);数据库分析结果示NEDD1与CDK2的表达呈正相关。结论:NEDD1具有抑制肺癌细胞凋亡、加速细胞周期进程从而促进细胞增殖的能力,且其在肺腺癌组织中的高表达与患者预后不良相关,因此可以作为肺腺癌诊断和预后新的标志分子。

Knock-down of NEDD1 expression inhibits lung adenocarcinoma cell proliferation and migration

AIM:To explore the role of neural precursor cell expression developmentally down-regulated protein 1(NEDD1)in the development and progression of lung cancer.METHODS:The differences of NEDD1 expression levels between lung cancer tissues and tumor-adjacent tissues were analyzed by the method of immunohistochemistry and TCGA database.Kaplan-Meier curve was used to analyze the correlation between lung cancer prognosis and the expression level of NEDD1.The proliferation of A549 cells was tested by plate colony formation experiment after knock-down of NEDD1 expression.The apoptosis rate and cell cycle distribution were examined by flow cytometry.The migration ability of the A549 cells was detected by Transwell assay.The protein levels of cell cycle-related molecules were determined by Western blot.Database analysis was performed to evaluate the relationship between the expression of NEDD1 and cyclin-dependent kinases 2(CDK2).RESULTS:Compared with the tumor-adjacent tissues,the expression level of NEDD1 in the lung cancer tissues was increased,so as the database analysis,and the higher expression of NEDD1 showed a poorer prognosis.Under light microscope,the A549 cells showed a low proliferation rate after silencing the NEDD1 expression,and the colony formation ability of the cells was also reduced;knock-down of NEDD1 expression induced the apoptosis and inhibited the cell migration;knock-down of NEDD1 expression blocked the cells in G1/S phase,and the protein levels of p-Rb and cyclinD1 were decreased,while the protein levels of p-Chk1,p-Chk2 and p-p53 were increased(P<0.05).A positive correlation between the expression of NEDD1 and CDK2 was noted by database analysis.CONCLUSION:NEDD1 plays an crucial role in promoting cell proliferation via inhibiting apoptosis and accelerating cell cycle,high expression of NEDD1 in lung adenocarcinoma tissue is related to poor prognosis,thus NEDD1 may be used as a candidate marker molecule for the diagnosis and prognosis of lung cancer.