慢病毒介导KAP-1增强胰腺癌细胞Capan-2侵袭能力  

Slow virus mediated KAP-1 effectively enhanced invasion of human pancreatic cancer Capan-2 cells

作　　者：江建新[1] 詹磊[1] 黄洋[1] 何燕浙[1] 喻超[1] 孙诚谊[1] 

机构地区：[1]贵阳医学院附属医院肝胆外科,贵州贵阳550004

出　　处：《中国现代医学杂志》2013年第13期16-19,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:81160311);贵州省肝胰疾病研究科技创新人才团队资助项目(No:黔科合人才团队[2010]4010);贵州省科教青年英才培养工程基金资助项目(No:黔省专合字[2012]177号);贵州省科技厅2011年度社会攻关计划基金资助项目(No:黔科合SY字[2011]3007)

摘　　要：目的观察KAP-1基因对人胰腺癌细胞Capan-2侵袭能力的影响,探求该基因作为治疗靶点的可行性。方法针对KAP-1基因构建真核表达载体,在293T细胞中包装成重组慢病毒Lv-eGFP-KAP-1,RT-qPCR和Western Blot法检测其对Capan-2细胞KAP-1的影响,Transwell侵袭小室检测其对Capan-2细胞侵袭能力的影响。结果成功地构建Lv-eGFP-KAP-1,感染Capan-2后细胞侵袭数目(128.3±4.6)较空白对照组(45.3±2.6)与阴性对照组(36.2±3.4)明显增加,差异有显著性(P<0.05)。结论 Lv-eGFP-KAP-1能显著上调Capan-2细胞KAP-1的表达,增强其侵袭能力,KAP-1基因有可能成为胰腺癌基因治疗的新靶点。【Objective】 To investigate the impact of KAP-1 on invasion of human pancreatic cancer Capan-2 cells,and to explore the feasibility of human KAP-1 gene as therapeutic target for pancreatic cancer.【Methods】 The KAP-1 cDNA was constructed to the pGC-LV plasmid and packed into the recombinant lentivirus Lv-eGFP-KAP-1 in 293T cells.The titer of lentivirus was determined by hole-by-dilution titer assay.The expression effect of Lv-eGFP-KAP-1 in Capan-2 cells was validated by real-time PCR and Western blot.After Capan-2 cells were infected with Lv-eGFP-KAP-1,cell invasion was detected by Transwell chamber assay.【Results】 Lv-eGFP-KAP-1 was successfully constructed.The titer of lentivirus was determined on 2×108 TU/mL.Overexpression of KAP-1 gene significantly enhanced invasion of human pancreatic cancer Capan-2 cells compared to blank control and negative control cells [（128.3±4.6） vs（45.3±2.6）,（36.2±3.4）,all P ＆lt;0.05）].【Conclusion】 Lv-eGFP-KAP-1 could effectively up-regulate the expression of KAP-1 gene in Capan-2 cells in vitro,significantly enhance cell invasion.KAP-1 might serve as a new target for gene therapy of pancreatic cancer.

关 键 词：胰腺癌 KAP-1 细胞侵袭 

分 类 号：R735.9[医药卫生—肿瘤]