Effect of AP-2α gene transfection on proliferation and invasion potential of human colon cancer SW480 cells in vitro
Ying Wáng, Xiaoqin Lan, Meining Li, Yuehong Zhang, Niuliang Cheng
Abstract
Ying Wáng, Xiaoqin Lan, Meining Li, Yuehong Zhang, Niuliang Cheng
Abstract
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. Key words: Colon neoplasms; Transcription factor AP-2; Gene transfection; SW480 cells; Cell proliferation; Invasiveness
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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. Key words: Colon neoplasms; Transcription factor AP-2; Gene transfection; SW480 cells; Cell proliferation; Invasiveness
Key concepts: Transfection, Molecular biology, Gentamicin protection assay, Cell growth, Biology, In vitro, Cell culture, Western blot