Establishment of a cellular model for Alzheimers disease by overezpressing Swedish-type mutated APP695
Objective:To establish a cellular model for investigating the relationship between Alzheimers disease and its risk factors. Methods: 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 800μg/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. Results: Restriction endonuclease digestion and sequencing analysis confirmed the authenticity of the eukaryotic expression vector pcDNA3.1-APP695sw; the production of Aβ1-40 and Aβ1-42 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. Conclusion: SH-SY5Y transfected with APP695sw can be used as the cellular model for the studies of pathogenesis and therapy in Alzheimers disease.
Alzheimers disease Cellular model Amyloid precursor peptide Swedish-type mutation Neuroblastoma cell
Tao Ma Wensheng Zhang
College of Resource Science & Technology,Beijing Normal University.Center for Natural Medicine Engineering (Beijing Normal University),Ministry of Education.Beijing,P.R.China
国际会议
北京
英文
1-4
2009-06-11（万方平台首次上网日期，不代表论文的发表时间）