Preparation of the Cells Transfected with Human APPswe695 Gene and the Determination of Aβ Production

Abstract: Alzheimer’s disease (AD) is a neurodegenerative disorder， and amyloid-β (Aβ) precursor protein (APP) is related to AD development. In this study， we developed a cell line that stably expressed APP gene with Swedish mutation (APPswe695 cells) and the production of amyloid-β was investigated. The expression plasmid with expressing APPswe695 gene was transfected into SH-SY5Y cells mediated by lentiviral vector. The gene expression of APPswe695 was confirmed at the levels of transcription and translation. The location of APP was analyzed by the method of immunocytochemistry. Aβ peptide， a product derived from APP by β-cleavage， was detected by the method of enzyme-linked immunosorbent assay (ELISA). The results indicated that APPswe695 cells could transcribe the mRNA and translate the protein of the transfected APPswe695 gene. The Swedish mutant protein of APP695 (APPswe695) had the same cellular distribution as endogenous APP770. Interestingly， APPswe cells are inclined to generate more intracellular but not extracellular Aβ. In general， these results indicated that APPswe695 cells can express the transfected APPswe695 gene and that APPswe695 is prone to generate more intracellular but not extracellular Aβ. This study implied that the APPswe695 cells can be used to investigate the pathogenesis and drug therapy of Alzheimer’s disease.

CSTR: 32200.14.cjcb.2019.01.0008