CBP基因对肺鳞癌NCI-H520细胞生长侵袭的抑制作用

目的:研究CSK结合蛋白(CSK-binding protein, CBP)基因转染对人肺鳞癌NCI-H520细胞体外生长侵袭的影响?方法:构建CBP真核表达质粒pcDNA3.0-CBP,以脂质体转染法转染体外培养的人肺鳞癌细胞株NCI-H520,G418筛选出抗性克隆?Western-blotting和RT-PCR分别检测转染前后CBP蛋白和mRNA水平的变化,MTT法分析细胞生长抑制作用,Transwell体外侵袭实验和Wound-healing实验观察细胞的侵袭和迁移能力?结果:稳定转染CBP基因的细胞株有外源目的基因的整合和相应蛋白的高表达?MTT检测表明,pcDNA3.0-CBP转染组活细胞数低于未转染组和pcDNA3.0(-)空载体质粒细胞转染组(P < 0.01)?细胞侵袭?迁移实验表明转染pcDNA3.0-CBP的瘤细胞侵袭与迁移能力均明显下降(P < 0.01)?结论:外源性CBP基因稳定转染可抑制人肺鳞癌NCI-H520细胞增殖?侵袭的恶性表型?

Growth and invasion inhibitory effects of CBP gene on lung squamous carcinoma cell line NCI-H520

Objective:To investigate the effects of exogenous wild CSK-binding protein(CBP) gene stably transfection on growth and invastion of lung squamous carcinoma cells in vitro. Methods:A recombinant eukaryotic expression plasmid pcDNA3.0-CBP was constructed. Human lung cancer cell line NCI-H520 was transfected with pcDNA3.0-CBP or mock transfected plasmid pcDNA3.0(-)with lipofectamine2000, and cells stably expressing CBP were screened out by G418(800 mg/L).Changes of CBP protein and mRNA levels were measured by Western blotting and RT-PCR, cell viability was tested by MTT assay. Transwell and wound-healing methods were used to detect the difference of invasion and migration between transfected and non-transfected cells. Results: CBP protein and mRNA levels in pcDNA3.0-CBP transfected NCI-H520 cells were significantly higher than those in control NCI-H520 cells and pcDNA3.0(-) transfected NCI-H520 cells. The MTT assay revealed that the CBP gene transfected cells had less proliferative ability than control cells and pcDNA3.0(-) transfected NCI-H520 cells(both P < 0.01). As compared with control cells and pcDNA3.0(-) transfected NCI-H520 cells, the invasion and migration ability of pcDNA3.0-CBP transfected cells decreased obviously(P < 0.01). Conclusion: The CBP gene can restrain malignant phenotypes of the human lung squamous carcinoma in vitro and may participate in construction of negative feedback loop of SFKS to inhibit growth, migration and invasion of lung cancer cells.