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目的:研究外源性CSK结合蛋白(CBP)基因转染对人肺腺癌SPC-A1细胞体外生长的影响。方法:构建CBP基因的真核表达质粒pcDNA3.0-CBP,应用重组质粒pcDNA3.0-CBP和空载体质粒pcDNA3.0(-),以脂质体转染法转染体外培养的人肺腺癌细胞株SPC-A1,用G418(800mg/L)筛选出抗性克隆。Western blot检测转染前后CBP蛋白水平的变化,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腺癌SPC-A1细胞增殖、侵袭的恶性表型。
Objective: To investigate the effect of exogenous CSK-binding protein (CBP) gene transfection on the growth of human lung adenocarcinoma SPC-A1 cells in vitro. Methods: The eukaryotic expression plasmid pcDNA3.0-CBP of CBP gene was constructed. The recombinant plasmid pcDNA3.0-CBP and empty vector plasmid pcDNA3.0 (-) were constructed and transfected into human lung adenocarcinoma Cancer cell line SPC-A1, with G418 (800mg / L) screened out resistant clones. The changes of CBP protein levels before and after transfection were detected by Western blot. The cell growth inhibition was analyzed by MTT assay. The invasion and migration of cells were observed by Transwell in vitro invasion assay and Wound-healing assay. Results: The cell lines stably transfected with CBP gene had the integration of exogenous gene and high expression of corresponding protein. MTT assay showed that the number of viable cells in pcDNA3.0-CBP transfection group was lower than that in non-transfection group and pcDNA3.0 (-) empty vector plasmid transfection group (P <0.01). Cell invasion and migration experiments showed that the invasion and migration ability of pcDNA3.0-CBP transfected cells were significantly decreased (P <0.01). Conclusion: The stable transfection of exogenous CBP gene can inhibit the malignant phenotype of human lung adenocarcinoma SPC-A1 cells in proliferation and invasion.