Bc1-2 基因加强表达对 OA 诱发的细胞编程死亡的效应

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．

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

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.