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奥替普拉和反义GST |
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柏华 胡国刚 朱明 魏慧娟 田海梅 罗贤懋
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pDORneo构建GST-π重组质粒pDGSTas，用Lipofectin将反义GST-πRNA导入转化的恶性Rat-1细胞(T-Rat-1)，经G418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 ELISA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130～140mg/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 mRNA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。
关键词：奥替普拉；凋亡；谷胱甘肽-硫-转移酶
分类号：R979.1 文献标识码：A
文章编号：0529-1356(2000)01-29
EFFECT OF OLTIPRAZ AND ANTISENCE GST-π ON
APOPTOSIS OF MALIGNANT TRANSFORMED CELLS
BAI Hua
(Cancer Institute,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100021,China)
HU Guo-gang
(Cancer Institute,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100021,China)
ZHU Ming
(Cancer Institute,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100021,China)
WEI Hui-juan
(Cancer Institute,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100021,China)
TIAN Hai-mei
(Cancer Institute,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100021,China)
LUO Xian-mao
(Cancer Institute,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100021,China)
Abstract：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 a[1] [2] [3] 下一页
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