AP-2α基因转染对人类结肠癌SW480细胞体外增生及侵袭能力的影响  

Effect of AP-2α gene transfection on proliferation and invasion potential of human colon cancer SW480 cells in vitro

作　　者：王莹[1] 兰小琴[1] 李美宁[1] 张悦红[1] 梁小波[2] 程牛亮[1] 

机构地区：[1]山西医科大学生物化学与分子生物学教研室,太原030001 [2]山西省肿瘤医院肛肠外科

出　　处：《肿瘤研究与临床》2008年第6期376-379,共4页Cancer Research and Clinic

基　　金：山西省科技攻关项目(2006031087-02)

摘　　要：目的 探讨AP-2α基因对人类结肠癌SW480细胞体外增生及侵袭能力的影响.方法 构建PODNA3.1(+)-AP-2α真核表达载体,利用脂质体介导pcDNA3.1(+)-AP-2α和pcDNA3.1(+)转染5W480细胞,并以正常SW480细胞作为空白对照;采用RT-PCR与Western blotting分别检测转染48 h后各组细胞中AP-2αmRNA与蛋白的表达情况;采用平板克隆、软琼脂克隆形成试验以及Transweil侵袭试验,分别检测各组细胞体外增生及侵袭能力.结果 SW480细胞内源性AP-2α蛋白表达缺失;转染AP-2α基因后,在SW480细胞中可检测到高水平的AP-2αmRNA及蛋白,细胞克隆形成率降低(P<0.05),软琼脂克隆体积小且数量少,细胞侵袭能力下降(P<0.05).结论 转染AP-2α基因可以抑制人类结肠癌SW480细胞的体外恶性增生以及侵袭能力.Objective To evaluate the proliferation and invasion potential of human colon cancer SW480 cells transfected by AP-2α gene in vitro. Methods pcDNA3.1 (+)-AP-2α was created by cloning AP-2α eDNA into the EcoRI site of poDNA3.1 (+). LipofectamineTM 2000 Reagent was used to mediate the transfection of pcDNA3.1(+)-AP-2α and pcDNA3.1 (+), and normal SW480 celia were cultured as a negative control. The mRNA and protein level of AP-2α in the cells of each group were detected 48 h after being transfected with plasmids above by RT-PCR and Western blotting analysis. The cell proliferation and invasion potential were examined by colony formation assay and Transwell invasion assay respectively. Results Lack of AP-2α protein expression in SW480 cells was verified by western blot analysis. After being transfected with AP-2α gene, the mRNA amount and protein expression increased dramatically, while the colony formation efficiency decreased(P <0.05), the cell proliferation in soft agar was inhibited, and the ability of its invasion dropped off(P <0.05) in vitro. Conclusion AP-2α gene suppresses the proliferation and invasion potential of human colon cancer SW480 cells in vitro.

关 键 词：结肠肿瘤 转录因子AP-2 基因转移 SW480细胞 细胞增殖 侵袭 

分 类 号：R735[医药卫生—肿瘤]