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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 explore the role of oltipraz and antisense GST-π in inducing apoptosis of malignant transformed cells. Methods The retroviral vector pDORneo was used to construct GST-π recombinant plasmid p DGSTas. Antisense GST-πRNA was transfected into transformed Rat-1 cells (T-Rat-1) by L ipofectin. Stained Rat-1 cells (AT-Rat-1); and then treated with oltipraz respectively against normal Rat-1, T-Rat- 1 and AT- Rat- 1; Gel electrophoresis analysis. Results ELISA analysis showed that the content of GST-πprotein in AT-Rat-1 cells was decreased compared with that in T-Rat-1 cells. After multiple doses were screened, the concentration of oltipraz in the range of 130-140 mg / Apoptosis. At the same time, Northern Blot examination showed that c-fos m RNA expression level of AT-Rat-1 cells was higher than that of normal Rat-1 cells and T-Rat-1 cells. Conclusion oltiprz can induce AT-Rat-1 cell apoptosis at the appropriate concentration. The low expression of GST-π and the high expression of c-fos gene are related to the apoptosis of AT-Rat-1 cells.