摘要
目的:研究ATRA联合IFN-α对人激素非依赖性前列腺癌PC-3细胞株的生长抑制作用及对凋亡相关基因GRIM-19和原癌基因STAT3表达的影响。方法:采用MTT比色法筛选ATRA和IFN-α联合应用的最佳时间和剂量;相差显微镜及吖啶橙染色观察其对PC-3细胞的促凋亡作用;DNA ladder提取试剂盒检测用药后细胞凋亡;RT-PCR方法观察用药后PC-3细胞GRIM-19 mRNA的表达;Western blot分别检测GRIM-19、STAT3的蛋白表达。结果:ATRA 1μmol/L联合IFN-α1 000 U/mL作用72 h对PC-3细胞有显著的增殖抑制作用,与溶媒对照组、ATRA或IFN-α单独应用组比较,差异显著(P<0.05)。形态学观察两药合用后PC-3细胞呈现典型的凋亡征象。DNA电泳图谱在ATRA和IFN-α两药合用组可见较AT-RA单独应用组更为明显的DNA ladder,IFN-α单独用药组及对照组没有出现DNA降解。RT-PCR结果表明,ATRA和IFN-α两药合用GRIM-19 mRNA表达增强,与对照组和ATRA、IFN-α组比较,差异显著(P<0.05)。Western blot结果显示,两药合用组GRIM-19蛋白表达增强,STAT3蛋白表达减弱,与对照组、ATRA、IFN-α组比较,差异有显著性(P<0.05)。结论:ATRA和IFN-α联合应用可抑制PC-3细胞的增殖并诱导凋亡,其机制之一可能是通过上调GRIM-19基因表达,进而下调原癌基因STAT3表达实现的。
Objective： To study the effects of trans retinoid acid （ATRA） combined with interferona（IFN-α） on the growth of androgen-independent prostate cancer PC-3 cell line and apoptosis-related gene GRIM-19 and oncogene STAT3 expression. Methods;PC-3 cells were treated with ATRT in combination or not with INF-α in vitro. Cell proliferation was evaluated by MTT assay, Cell morphological changes were observed under phase-contrast microscope. Apoptotie cells were detected by AO staining and DNA ladder agarose gel electrophoresis. GRIM-19 mRNA expression was measured by RT-PCR. GRIM-19 and STAT3 protein expression was determined by Western blot. Results： Compared with control, IFN-α,or ATRA group, proliferation of PC-3 cells was significantly inhibited after incubation with ATRA 1 /Lmol/L plus IFN-α 1 000 IU/mL for 72 h（P〈0.05）. Cells showed apoptotic characteristics after ATRA plus IFN-α treatment. DNA was degraded more in ATRA plus IFN-α group than that in ATRA group. No DNA degradation was observed in IFN-α group and control group. GRIM-19 mRNA and protein expression was increased but STAT3 protein expression was decreased after ATRA plus IFN-α treatment （P〈0. 05）. Conclusion： ATRA in combination with INF-α suppressed the proliferation and induced apoptosis of PC-3 cells partly by up-regulation of GRIM-19 gene expression and down-regulation of STAT 3.
出处
《肿瘤》
CAS
CSCD
北大核心
2006年第3期228-231,共4页
Tumor
基金
国家科技部国际科技合作重点项目(编号:2004DFB02000)
中日政府间专项技术合作项目(编号:59)
