【摘要】 目的:观察上调核型Clusterin(nuclear clusterin,nCLU)的表达对人肺腺癌A549细胞生物学行为的影响。方法:构建插入核型Clusterin基因的真核表达质粒,采用脂质体法转染A549细胞,潮霉素B筛选得到稳定转染的细胞株以及对照细胞株;Western印迹法检测转染前后A549细胞中Clusterin蛋白的表达;CCK-8法检测被转染细胞体外增殖的改变;FCM法检测细胞周期分布的改变;体外迁移和侵袭实验检测细胞体外运动迁移和侵袭能力的变化。结果:nCLU表达的上调,可使A549细胞体外增殖能力明显减慢;G0/G1期细胞比例明显增加,由原来的(33.54±2.10)%上升到(63.31±4.30)%;体外迁移及侵袭能力明显减弱(P<0.05)。结论:上调nCLU的表达可明显抑制肿瘤细胞的增殖,降低细胞的运动迁移和侵袭能力。更多还原

【Abstract】 Objective:To observe the up-regulation of nuclear Clusterin (nCLU)gene on the biological behaviors of human non-small cell lung cancer cell line A549.Methods:Sense eukaryotic expression vector of nCLU was constructed by cloning the cDNA of nCLU into pIREShyg3 vector. A549 cells were transfected with pIRES-nCLU and pIREShyg3 vectors by lipofectAMINETM 2000 mediation,respectively. Stable transfected cells were selected by hygromycin B screening. CCK-8 assay was performed to evaluate the effect of nCLU over-expression on cell proliferation in vitro. The expression level of nGLU protein was examined by Western blotting. Cell cycle distribution was detected by FCM with PI staining. The alteration of migration and metastasis potential of A549 cells before and after nCLU gene transfection were assayed by cell chemotactic migration and invasion test. Results:The proliferation speed of the transfected A549 cell clones stably over-expressing nCLU was slowed down. FCM analyses revealed that the percentage of cells in G0/G1 phase dramatically increased from (33.54±2.10)% to (63.31±4.30)%. The cell chemotactic migration and invasion potentials were markedly reduced after nCLU gene transfection (P<0.05). Conclusion:Up-regulation of nCLU can greatly inhibited the proliferation and decreased the migration and invasion capabilities of A549 cells.更多还原