慢病毒导入的CDC25B2 siRNA对胰腺癌细胞CFPAC-1生物学行为的影响  

Effects of lentivirai-delivered CDC25B2 siRNA on biological behaviors of human pancreatic cancer cell line CFPAC-1

作　　者：杨正平[1] 石欣[1] 肖志[1] 张齐[1] 孔波[1] 严伟[1] 葛梓[1] 

机构地区：[1]东南大学附属中大医院普外科,南京210009

出　　处：《国际肿瘤学杂志》2008年第10期792-797,共6页Journal of International Oncology

基　　金：国家自然科学基金资助项目（30500491）

摘　　要：目的利用已构建成功的、在人胰腺癌细胞株CFPAC-1中稳定抑制细胞分裂周期25同源体B（CDC2582）的重组慢病毒，建立CDC2582表达降低的细胞株，以研究该基因的作用。方法将产生CDC2582小干扰RNA（siRNA）分子的、携带绿色荧光蛋白的重组慢病毒感染CFPAC-1细胞后，用流式细胞仪筛选稳定表达的细胞株。应用RT—PCR和Westernblot分别对细胞中CDC2582的mRNA和蛋白表达水平进行分析。应用MTT法、流式细胞仪、平板克隆形成和Transwell小室法分别检测其对细胞增殖、周期、细胞克隆形成能力和迁移侵袭能力的影响。结果CDC2582 siRNA显著下调CFPAC-1细胞中CDC2582的表达，所构建的重组慢病毒导入的siRNA有较高的沉默效率，CFPAC-1细胞的增殖能力、克隆形成能力以及迁移侵袭能力显著增强，细胞周期无明显改变。结论CDC2582基因可显著影响胰腺癌CFPAC-1细胞的增殖、克隆形成和迁移能力，为利用CDC2582所介导的信号通路进行肿瘤基因治疗提供了实验依据。Objective To establish CFPAC-1 cell lines deficient in CDC25B2 by recombinant lentivirus, and to investigate the role of this gene. Methods After CFPAC-1 cells were transduced with recombinant lentivirus producing CDC25B2 siRNA, stably transduced cells with green fluorescent protein were selected by flow cytometer. The mRNA and protein expression of CDC25B2 was examined by RT-PCR and Western blot analysis. The effect of the lentivirus on the cell proliferation, cell cycle, clone-forming, migration and invasion ability was analyzed by MTT method, flow cytometer, plate clone-forming assay and Transwell chamber method respectively. Results CDC25B2 siRNA knocked down CDC25B2 expression in CFPAC-1 cells significantly. The silencing efficiency of siRNA transduction by recombinant lentivirus was very high. Proliferation, cloneforming, migration and invasion ability of human pancreatic cancer cell line CFPAC-1 were significantly increased, while cell cycle was not affected. Conclusion CDC25B2 plays an important role in cell proliferation, clone-forming, migration and invasion of pancreatic cancer. This research provides experimental evidences for targeting CDC25B2 in gene therapy against pancreatic cancer.

关 键 词：基因 cdc 胰腺肿瘤 慢病毒属 RNA 小分子干扰 细胞增殖 

分 类 号：R735[医药卫生—肿瘤]