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目的:探讨核蛋白1(Nupr1)调控非小细胞肺癌细胞迁移、凋亡机制的研究。方法:肿瘤抑制剂盐酸素(salinomycin)不同时间处理非小细胞肺癌细胞A549后采用Western Blot法检测非小细胞肺癌细胞A549中Cleaved Caspase-3、Nupr1的蛋白表达;Transwell小室检测Nupr1基因沉默后非小细胞肺癌细胞A549细胞体外迁移、侵袭能力的变化;Western Blot法检测Nupr1沉默后非小细胞肺癌细胞A549 MMP-2、TIMP-1的蛋白表达;流式细胞仪检测Nupr1沉默后非小细胞肺癌细胞A549的凋亡情况。结果:与未经肿瘤抑制剂salinomycin处理对照组相比较,salinomycin处理后的非小细胞肺癌细胞A549中Nupr1蛋白表达量下降,Cleaved Caspase-3蛋白表达量升高,并且随着作用时间呈依赖关系。Nupr1-siRNA转染组的迁移能力相比对照组未转染组下降(64.4±7.2)%,Nupr1-siRNA转染组的侵袭能力相比对照组下降(58.7±7.3)%。与未转染Nupr1-siRNA对照组相比较,转染后TIMP-1的表达明显上调,而MMP-2的表达则明显下调。流式细胞仪检测结果显示Nupr1沉默后非小细胞肺癌细胞A549出现大量凋亡。结论:Nupr1基因沉默后通过上调TIMP-1的表达,下调MMP-2的表达降低肺癌A549细胞的侵袭和迁移能力,进而促进非小细胞肺癌细胞凋亡。
Objective: To investigate the mechanism of Nupr1 in regulating the migration and apoptosis of non-small cell lung cancer cells. Methods: The expression of Cleaved Caspase-3 and Nupr1 in non-small cell lung cancer A549 cells was detected by Western Blot after treated with tumor suppressor salinomycin for different time. Transwell chamber was used to detect the expression of Nupr1 gene in non-small cell lung cancer cells after being silenced Small cell lung cancer A549 cells in vitro migration and invasion ability changes; Western Blot detection of Nupr1 silencing non-small cell lung cancer A549 MMP-2, TIMP-1 protein expression; Flow cytometry Nupr1 silent non-small cell lung cancer Apoptosis of A549 cells. RESULTS: Compared with the untreated salinomycin control group, Nupr1 protein expression and Cleaved Caspase-3 protein expression in salinomycin-treated non-small cell lung cancer A549 cells increased, and with the time dependent . The migration ability of Nupr1-siRNA transfection group was decreased (64.4 ± 7.2)% compared with untransfected control group, and the invasion ability of Nupr1-siRNA transfection group was (58.7 ± 7.3)% lower than that of control group. Compared with untransfected Nupr1-siRNA control group, the expression of TIMP-1 was up-regulated and the expression of MMP-2 was down-regulated after transfection. Flow cytometry results showed that Nupr1 silenced a large number of apoptosis in non-small cell lung cancer A549 cells. CONCLUSIONS: Nupr1 silencing can up-regulate the expression of TIMP-1, down-regulate the expression of MMP-2 and decrease the invasion and migration of lung cancer A549 cells, and then promote the apoptosis of non-small cell lung cancer cells.