CAJ | 학술논문

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.
목적 탐토AP-2α기인대인류결장암SW480세포체외증생급침습능력적영향.방법 구건PODNA3.1(+)-AP-2α진핵표체재체,이용지질체개도pcDNA3.1(+)-AP-2α화pcDNA3.1(+)전염5W480세포,병이정상SW480세포작위공백대조;채용RT-PCR여Western blotting분별검측전염48 h후각조세포중AP-2αmRNA여단백적표체정황;채용평판극륭、연경지극륭형성시험이급Transweil침습시험,분별검측각조세포체외증생급침습능력.결과 SW480세포내원성AP-2α단백표체결실;전염AP-2α기인후,재SW480세포중가검측도고수평적AP-2αmRNA급단백,세포극륭형성솔강저(P<0.05),연경지극륭체적소차수량소,세포침습능력하강(P<0.05).결론 전염AP-2α기인가이억제인류결장암SW480세포적체외악성증생이급침습능력.
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.