Bc1-2 基因加强表达对 OA 诱发的细胞编程死亡的效应

Effect of Enforced Bcl 2 Over expression on OA induced Cell Death in Human Neuroblastoma SK Cell Line

ＯＡ能诱发人神经母细胞瘤ＳＫ细胞进行编程死亡．死亡细胞缩小变圆，细胞质凝聚，ＤＮＡ有控降解成约２００ｂｐ左右的片段，且这一过程可由蛋白质合成抑制剂亚胺环己酮（ＣＨＸ）抑制．将编码Ｂｃ１－２全长蛋白质的ｃＤＮＡ植入ｐＸＪ４１ｎｅｏ载体中，使其表达由ＨＣＭＶ病毒启动子控制．形成的顺义（ｐＢｃｌ－２－Ｓ）及反义（ｐＢｃｌ－２－ＡＳ）表达质粒经转染导入ＳＫ细胞中获得稳定转染子，Ｗｅｓｔ－ｅｒｎ印迹表明顺义转染子表达较大量的２６ｋｄＢｃ１－２蛋白，而反义转染子则不表达．增强表达的Ｂｃ１－２基因产物对ＯＡ引ＳＫ细胞编程死亡无抑制效应．

OA induce cell death in the human neuroblastoma SK cell line with the characteristic feather of apoptosis.Cells are rounded and shrink;cytoplasm condenses;DNA is fragmented into a 200 bp ladder.Moreover,the above phenomenon can be partially inhibited by protein synthesis inhibitor cycloheximide.A 1.6 kb NDA encoding the full length human Bcl 2 protein was inserted into mammalian expression vector pXJ41neo and placed under the control of HMCV promoter in either the sense (pBcl 2 S) or antisense (pBcl 2 As) orientation.The above two plasmids were transfected into SK cell and stable transfectants were obtained.pBcl 2 s transfected Sk cells expressed high level of 26 kd Bcl 2 protein,as evidenced by Western blotting,while pBcl1 2 As transfected cells showed no detectable Bcl 2 expression.Over expressed Bcl 2 protein cannot inhibit OA induced cell death.