奥替普拉和反义GST-π对恶性转化细胞凋亡影响的研究  

EFFECT OF OLTIPRAZ AND ANTISENCE GST-π ON APOPTOSIS OF MALIGNANT TRANSFORMED CELLS

作　　者：柏华[1] 胡国刚[1] 朱明 魏慧娟[1] 田海梅[1] 罗贤懋[1] 

机构地区：[1]中国协和医科大学中国医学科学院肿瘤研究所,北京100021

出　　处：《解剖学报》2000年第1期29-33,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金!( 3 95 70 798)

摘　　要：目的　探索奥替普拉 (oltipraz)和反义 GST- π在诱导恶性转化细胞凋亡中的作用。　方法　利用逆转录病毒载体 p DORneo构建 GST- π重组质粒 p DGSTas,用 L ipofectin将反义 GST- πRNA导入转化的恶性 Rat- 1细胞 (T- Rat- 1) ,经 G41 8筛选获得反义转染的 Rat- 1细胞 (AT- Rat- 1) ;再用 oltipraz分别作用于正常的 Rat- 1、T-Rat- 1和 AT- Rat- 1;细胞凋亡状况用流式细胞仪和 DNA凝胶电泳分析。　结果　 EL ISA分析显示 AT- Rat- 1细胞的 GST- π蛋白含量比 T- Rat- 1细胞降低 ;筛选多个剂量后发现 oltipraz在 130～ 140 mg/ L,范围能明显诱导 AT-Rat- 1细胞凋亡。同时用 Northern Blot检查发现 AT- Rat- 1细胞的 c- fos m RNA表达水平高于正常 Rat- 1细胞和T- Rat- 1细胞。　结论　 oltiprz在适当浓度时能诱导 AT- Rat- 1细胞凋亡 ;GST- π的低表达和 c- fos基因的高表达与 AT- Rat- 1细胞凋亡相关。Objective\ To Study the effect of oltipraz and anti sence GST π on apoptosis of malignant transformed Rat 1 cell Methods GST π cDNA inserted retroviral expression vectors pDORneo were constructed into recombinant pDGSTas and transfected to T Rat 1 cell by Lipofectin;T Rat 1 cell containing anti sense GST π RNA(AT Rat 1) was obtained by G418 screening; then Rat 1,T Rat 1 and AT Rat 1 cells were treated by oltipraz, The apoptosis of above cells was analyzed by DNA agarose gel electrophoresis and flow cytometry examination. Results GST π protein content of AT Rat 1 is lower than that of T Rat 1 by ELISA.Oltipraz has a obvious effect of inducing AT Rat 1 cell apoptosis in 130 140mg/L. Meanwhile, it is shown that c fos mRNA of AT Rat 1 cell is higher experssion than that of T Rat 1 by Northern blot analysis. Conclusion The apoptosis of AT Rat 1 cell can be induced by a effective dose of oltipraz; The high expression of c fos gene and low expression of GST π gene are related to AT Tat 1 cell apoptosis.

关 键 词：奥替普拉 肺肿瘤 GST-Π 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤] R979.1[医药卫生—临床医学]