【摘要】 目的本研究以人胃癌SGC-7901细胞为肿瘤模型,在体外探讨维生素E琥珀酸酯(VES)在抑制肿瘤细胞生长、诱导细胞凋亡过程中对NF-κB组成性激活及抗凋亡基因在转录水平上的调控作用。方法采用MTT法和AnnexinV法分别检测不同浓度水平VES处理人胃癌SGC-7901细胞后细胞的生长和凋亡情况。采用细胞成分分离法获得细胞核总蛋白,然后以Westernblot法测定VES处理后核因子蛋白p65在细胞核总蛋白中的含量。用RT-PCR方法检测VES对SGC-7901细胞中抗凋亡基因c-IAP1、c-IAP-2、NAPI、survivin、XIAP的转录调节作用。结果12.5～50.0μmolVES作用24h后,SGC-7901细胞的生长被明显抑制。50μmolVES作用于细胞12h后可诱导细胞发生凋亡,且作用48h后其凋亡诱导作用更明显。Westernblot结果发现,VES处理SGC-7901细胞后使细胞核内NF-κBp65蛋白的含量下降,即抑制了NF-κBp65向细胞核内的迁移。RT-PCR结果提示,VES处理胃癌细胞后可以在基因水平上抑制抗凋亡基因NAIP和survivin转录,对其他IAPs家族成员-c-IAP1、c-IAP-2、XIAP的转录无下调作用。结论VES对SGC-7901细胞内NF-κB组成性激活的调控作用及对抗凋亡基因在转录水平上的抑制作用可能是其抑制SGC-7901细胞生长、诱导细胞发生凋亡的机制之一。更多还原

【Abstract】 Objective To explore whether the regulation of NF-κB signaling pathways were involved in the anti-cancer effects of vitamin E succinate (VES) on human gastric carcinoma SGC-7901 cells. Method MTT assay, Annexin-V methods were used to detect the growth inhibition and apoptosis-inducing effects of VES on SGC-7901 cells. Western blot was applied to detect the protein expression of NF-κBp65 in cell nuclei and cell plasma. RT-PCR method was performed to investigate the NF-κB downstream genes expression, including c-IAP1, c-IAP2, NAIP, XIAP, and survivin. Results The growth of SGC-7901 cells was significantly inhibited in dose-dependent manner after 12.5-50μM VES treatment for 24h. 50μM VES induced apoptosis after treating cells for 12h, and the apoptosis-inducing effect was more distinctive at 48h. Western blot results showed that VES downregulated the protein expression of NF-κBp65 in cell nuclei accompanied with downregulation of NF-κB downstream anti-apoptosis gene expression of survivin and NAIP without changing the gene expression of c-IAP1, c-IAP2 and XIAP. Conclusion VES inhibited the growth and induced apoptosis of SGC-7901 cells with inhibition of NF-κBp65 constitutive activity, which was probably contributed to VES-mediated downregulation of anti-apoptosis gene expression in human gastric carcinoma cells.更多还原