Nupr1调控非小细胞肺癌细胞迁移、凋亡机制的研究

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。

Nupr1 Regulates the Migration and Apoptosis of Non-small Cell Lung Cancer Cells

ABSTRACT Objective: To explore the migration and apoptosis mechanism of Nupr1 in non-small cell lung cancer cells (NSCLC). Methods: Western Blot was used to detect the protein expression of Cleaved Caspase-3 and Nupr1 in NSCLC A549 cells after treated by tumor suppressor salinomycin for different time. The Transwell inserts was applied to detect the changes of migration and invasion ability of A549 cells after silence of Nupr1. The MMP- and TIMP-1 protein expression in A549 cells were detected by western blot after Nupr1 gene silencing. The apoptosis of A549 cells after Nupr1 silencing was detected by flow cytometry. Results: Compared with control group which had no tumor suppressor salinomycin treatment, the Nupr1 protein expression was decreased in non-small cell lung cancer A549 cells which were treated by salinomycin, and the Cleaved Caspase-3 protein expression was increased in a time-dependent way. Migra- tion of Nupr1-siRNA group decreased by (64.4±7.2)%, compared with control group, and the invasion ability of Nupr1-siRNA group decreased by (58.7±7.3)%. The TIMP-1 expression was significantly up-regulated and the MMP-2 expression was significantly down-regulated as compared with control group. Flow cytometry results showed that NSCLC A549 cells were largely apoptotic after Nupr1 silencing. Conclusion: The inhibition of Nupr1 gene expression can reduce the invasion and migration of A549 cells co that can accelerate the apoptosis of NSCLC A549 cells by up-regulating the expression of TIMP-1 and down-regulating the expression of MMP-2.