CD基因对人胰腺癌细胞株Patu 8988杀伤作用的研究  

Killing Effect of Adenoviral Mediated Cytosine Deaminase Gene on Human Pancreatic Cancer Cell Line Patu 8988

作　　者：潘雪[1] 李兆申[1] 许国铭[1] 崔龙[1] 戴观荣[1] 屠振兴[1] 

机构地区：[1]第二军医大学长海医院,200433

出　　处：《实用癌症杂志》2002年第2期119-121,共3页The Practical Journal of Cancer

基　　金：上海科委重点课题资助 (No :9941 1 90 4 4 )

摘　　要：目的　探讨腺病毒介导胞嘧啶脱氨酶 (CD )基因对人胰腺癌细胞株Patu 8988的杀伤作用。方法　采用细菌内同源重组方法 ,构建含胞嘧啶脱氨酶 (CD)基因重组腺病毒载体 ,经 2 93细胞包装、扩增 ,氯化铯密度梯度离心制备纯化高效的重组腺病毒 ,体外转染人胰腺癌细胞株Patu 8988,并给予前药 5 Fc ,观察其体外杀伤效果。结果　含CD基因的腺病毒载体经酶切鉴定正确 ,包装纯化后 ,检测病毒滴度为 2× 10 11pfu/ml ,将重组腺病毒转染胰腺癌细胞株Patu 8988,加用前药 5 Fc后 ,转染CD基因的Patu 8988细胞及混育的胰腺癌细胞生长明显受抑制。结论　CD腺病毒不仅转染效果强 ,而且可直接或通过旁观者效应杀伤胰腺癌细胞。Objective To evaluate killing effects of cytosine deaminase gene mediated by adenovirus vector on human pancreatic cancer cell line Patu 8988.Methods Cytosine Deaminase(CD) gene was cloned into pAdTrack CMV CD,then pAdTrack CMV CD and pAdEasy 1 were recombinated in bacteria.The newly recombinated Ad CD containing green fluorescent protein (GFP) were propagated in 293 cells and purified by cesium chloride gradient centrifugation.Human pancreatic carcinoma cell line 8988 were infected with this virus,then 5 Fc was added,XTT assay was used to estimate relative numbers of viable cells.Results The positive clones were selected by endonuclease to digest the combinatants and the titre of virus containing CD gene was 2×1011 pfu/ml.It was found that significant cytotoxic activities were possessed by 5 Fc for CD gene transduced 8988 cell line,but little effects on the nontransduced pancreatic carcinoma cells.Conclusion CD gene mediated by adenovirus has a high infectivity and is efficient for killing cultured pancreatic carcinoma cells.These data demonstrate utilization of suicide gene prodrug stratege may be effective for combating pancreatic cancer.

关 键 词：胰腺癌 腺病毒载体 胞嘧啶脱氨酶 基因治疗 

分 类 号：R735.9[医药卫生—肿瘤]