Establishment of a Cellular Model for Alzheimer´s Disease by Overexpressing Swedish-Type Mutated APP695

The objective of this study is to establish a cellular model for investigating the relationship between Alzheimer´s disease and it´s risk factors. The full length cDNA fragment of Swedish-type mutated APP695 (APP695sw) was amplified by PCR and ligated into pcDNA3.1(+) vector to construct the recombinant plasmid pcDNA3.1-APP695sw. The SH-SY5Y neuroblastoma cells were transfected with pcDNA3.1-APP695sw by liposome-mediated method, and stable transfectants were selected in 800 mug/ml G418. Gene expression was examined with RT-PCR and Sandwich ELISA. The effects of APP695sw overexpression on SH-SY5Y were tested by growth curve assay and morphologic observation. Cell viability was measured with MTT assay at different time points. Restriction endonuclease digestion and sequencing analysis confirmed the authenticity of the eukaryotic expression vector pcDNA3.1-APP695sw; the production of Abeta1-40 and Abeta1-42 were remarkably enhanced in the pcDNA3.1-APP695sw transfected cells; there is no effect of APP695sw overexpression on cell morphology, meanwhile cell growth was postponed after APP695sw transfection. SH-SY5Y transfected with APP695sw can be used as the cellular model for the studies of pathogenesis and therapy in Alzheimer´s disease.