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作 者:温龙平[1] 蔡毓[1] 陈亚兵[1] 俞春东[1] 曾桂生[1]
机构地区:[1]厦门大学肿瘤细胞工程国家专业实验室
出 处:《厦门大学学报(自然科学版)》1997年第1期110-114,共5页Journal of Xiamen University:Natural Science
基 金:国家自然科学基金
摘 要: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.
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