摘要
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 effects of exogenous wild csk-binding protein（CBP）gene stably transfection on growth of lung adencarcinoma cells in vitro. Methods：at first,a recombinant eukaryotic expression plasmid pcDNA3.0-CBP was constructed. Human lung cancer cell line SPC-A1 was transfected with pcDNA3.0-CBP or mock transfected plasmid pcDNA3.0（-）with lipofectamine 2000,and cells that can express CBP stably were screened out by g418（800 mg/l）. Change of CBP protein level was measured by western-blot;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 level in pcDNA3.0-CBP transfected SPC-A1 cells was significantly higher than that in control SPC-A1 cells and pcDNA3.0 （-）transfected SPC-A1 cells. The mtt assay revealed that the CBP gene transfected cells had less proliferation ability than control cells and pcDNA3.0（-）transfected SPC-A1 cells （p 〈 0.01）. As compared with control cells and pcDNA3.0（-）transfected SPC-A1 cells,the invasion and migration ability of pcDNA3.0-CBP transfected cells decreased obviously（p 〈 0.01）. Conclusion：the CBP gene,as a recently identified transmembrane protein,can restrain malignant phenotypes of the human lung adencarcinoma in vitro and may participate in construction of negative feedback loop of sfks to inhibit lung cancer cells from growth,migration and invasion.
Keywords： csk-binding protein（CBP） human lung adencarcinoma transfection invasion
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第9期1204-1207,1223,共5页
Journal of Nanjing Medical University(Natural Sciences)
关键词
CBP基因
肺腺癌
转染
侵袭
CSK-binding protein（CBP）
human lung adencarcinoma
transfection
invasion
