摘要
目的 研究 MDM2癌基因与野生型 p5 3基因在细胞凋亡发生中的相互关系。方法 构建了一个表达人野生型 ( wt) p5 3逆转录病毒载体 ,经 PA31 7细胞包装后转染人胰腺癌细胞 ( PC- 2 ) ,建成 PC- 2 /swtp5 3转化细胞系。再用含 MDM2基因的p CMV- MDM2载体转染 PC- 2 /swtp5 3细胞 ,获得双重转染细胞系 PC- 2 /swtp5 3/p CMV- MDM2。用流式细胞技术 ,原位检测分析和 DNA凝胶电泳分析细胞凋亡。结果 恢复 wtp5 3表达的胰腺癌细胞系 ( PC- 2 /swtp5 3)细胞凋亡明显增多 ( >1 2 % ) ,而双重转染的细胞系 ( PC- 2 /swtp5 3/p CMV- MDM2 )则凋亡细胞数明显下降 ( 3.2 % )。结论 MDM2癌基因可抑制由野生型 p5 3基因诱发的细胞凋亡。
Objective To investigate the interaction between Murine Doube Minute2 ( MDM2 ) oncogene and p5 3gene in the evolution of apoptosis in pancreatic carcinoma.Methods A recombinant retroviral vector expressing wild- type p5 3was constructed and packaged bypackaging cell line PA31 7cells using calcium phosphate coprecipitation method.The viral su- pernatant of the packaged vector was used to transfect the pancreatic carcinoma cell line( PC- 2 ) ,a transformantcell line PC- 2 / swtp5 3was then established.A recombinant vector p CMV- MDM2 was transduced into PC- 2 / swtp5 3cells by lipofectin- mediated method,a double transfected cell line PC- 2 / swtp5 3/ p CMV- MDM2 was formed.Results By means of flow cy- tometry,in situ Td T analysis and DNA agarose gel electrophoresis,an increase of apoptosis was demonstrated in PC- 2 / swtp5 3cell line( >1 2 % ) ,whereas apoptosis was decreased in PC- 2 / swtp5 3/ p CMV- MDM2 cell line( 3.2 % ) .Conclusions MDM2 oncogene can inhibit apoptosis induced by wild- type p5 3gene in pancreatic carcinoma.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1998年第6期401-406,共6页
Acta Academiae Medicinae Sinicae
基金
国家教委博士学科点专项科研基金!( 960 0 2 3 2 7)
