人淀粉样前体蛋白基因C端在SH-SY5Y细胞系中的稳定表达及其对细胞生长发育的影响

The Stable Expression of Human APP Gene C terminal Fragment and Its Influence on the Growth and Development of SH SY5Y Cells

淀粉样前体蛋白（ＡＰＰ）是阿尔茨海默氏病（ＡＤ）病因学中重要的分子，但其正常的生理功能尚不清楚．为了研究细胞内过量产生其各种片段对细胞生理机能的影响．将人ＡＰＰ６９５ｃＤＮＡ中编码Ｃ端１０５个氨基酸残基的ＤＮＡ片段重组到真核表达载体ｐＤＯＲｎｅｏ中形成重组质粒ｐＤＯＲ－ｎｅｏ－ＣＴ．然后用脂质体将其转染到人神经母细胞瘤细胞ＳＨ－ＳＹ５Ｙ中．用８００μｇ／ｍｌＧ４１８筛选获得了在ｍＲＮＡ和蛋白质水平均表达相应片段的稳定细胞系．ＭＴＴ和ＬＤＨ分析表明，ＡＰＰＣ端的表达未能对ＳＨ－ＳＹ５Ｙ细胞产生明显的毒性作用．

The amyloid precursor protein (APP) is a molecule centrally involved in Alzheimer disease pathology.But its normal function is poorly understood.To investigate the consequences of increased intracellular production of various regions of APP on cellular physiology,the C terminal 105 amino acids of the human APP was inserted into retrovirus eukaryocytic expressing vector pDORneo to form recombinant pDORneo CT.Lipofectin was used to transfect the recombinant into human neuroblastome cell SH SY5Y.RT PCR and Western blot showed that stable transfectants which expressed the corresponding fragment of APP gene in mRNA and protein level had been obtained.Morphological observation and MTT,LDH assay showed that the expression of APP C105 resulted in no obvious toxic effect on transformed SH SY5Y cells.