Atox1参与调控肺癌细胞的增殖和迁移

Effect of Atox1 on Proliferation and Migration of Lung Cancer Cells

目的:研究Atox1基因对肺癌细胞增殖和迁移的影响。方法:通过UALCAN数据库在线分析Atox1在肺癌组织中的表达量;利用qRT-PCR和蛋白印迹检测转移性肺癌细胞系(NCI-H292、95-D)和非转移性肺癌细胞系(A549)中Atox1的mRNA和蛋白相对表达量;在NCI-H292细胞中敲低Atox1,分别利用CCK-8和Transwell检测NCI-H292细胞增殖和迁移能力;在A549细胞中过表达Atox1,检测细胞增殖和迁移能力;建立稳定敲降Atox1的NCI-H292细胞株,回复表达野生型或缺失核定位信号的突变型Atox1,分析Atox1功能与其亚细胞定位间的联系。结果:在线分析Atox1在肺癌组织中表达量明显高于癌旁组织;转移性肺癌细胞系(NCI-H292、95-D)中Atox1的mRNA和蛋白表达明显高于非转移性肺癌细胞系(A549);敲低Atox1能够抑制NCI-H292细胞增殖和迁移能力;过表达Atox1能够促进细胞A549细胞的增殖和迁移能力;缺失核定位信号的Atox1促肺癌细胞增殖和迁移的能力显著减弱。结论:Atox1促进肺癌细胞增殖和迁移能力,并且与其核定位和转录活性密切相关。

Objective:To investigate the effect of Atox1 gene on proliferation and migration of lung cancer cells.Methods:The expression of Atox1 in lung cancer tissues was analyzed online using UALCAN database.The relative expression levels of Atox1 mRNA and protein in metastatic lung cancer cell lines(NCI-H292,95-D)and non-metastatic lung cancer cell lines(A549)were measured by qRT-PCR and Western blotting.Atox1 was knocked down in NCI-H292 cells,and proliferation and migration of NCI-H292 cells were detected by CCK-8 and Transwell,respectively.Atox1 was overexpressed in A549 cells to detect the proliferation and migration of A549 cells.NCI-H292 cell line with stable Atox1 knockdown was established to restore the expression of wildtype or mutant Atox1 without nuclear localization signal,and the relationship between Atox1 function and its subcellular localization was analyzed.Results:The Atox1 expression in lung cancer tissues from online analysis was significantly higher than that in adjacent tissues.The mRNA and protein expression of Atox1 of metastatic lung cancer cell lines(NCI-H292,95-D)was significantly higher than that of non-metastatic lung cancer cell line(A549).Atox1 knocked down inhibited NCI-H292 cells proliferation and migration;overexpression of Atox1 promoted A549 cells proliferation and migration.The ability of Atox1 without nuclear localization signal to promote proliferation and migration of lung cancer cells was significantly reduced.Conclusion:Atox1 promotes the proliferation and migration of lung cancer cells,which is closely related to its nuclear localization and transcriptional activity.