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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。
OBJECTIVE: To investigate the effect of vitamin E succinate (VES) on the constitutive activation of NF-κB and the anti-apoptotic gene in the process of inhibiting tumor cell growth and inducing apoptosis in SGC-7901 human gastric cancer cell model Transcriptional regulation. Methods MTT and AnnexinV were used to detect the growth and apoptosis of human gastric cancer cell line SGC-7901 after treatment with different concentrations of VES. Cell total nuclear protein was obtained by cell fractionation method. Then the content of nuclear factor p65 in nucleus total protein was determined by Western blot. The transcriptional regulation of anti-apoptotic gene c-IAP1, c-IAP-2, NAPI, survivin and XIAP in SGC-7901 cells was detected by RT-PCR. Results The growth of SGC-7901 cells was significantly inhibited after treated with 12.5-50.0 μmol VES for 24 h. After treated with 50μmol VES for 12h, the cells could induce apoptosis, and the apoptosis induction effect was more obvious after 48h. Western blot results showed that the VES treated SGC-7901 cells decreased the content of NF-κB p65 in the nucleus, which inhibited the migration of NF-κB p65 into the nucleus. The results of RT-PCR indicated that VES could inhibit the transcription of anti-apoptotic genes NAIP and survivin at the gene level and had no effect on the transcription of other IAPs family members -c-IAP1, c-IAP-2 and XIAP. Conclusions The regulation of VES on the constitutive activation of NF-κB in SGC-7901 cells and the anti-apoptotic gene transcription at the transcriptional level may be one of the mechanisms by which VES inhibits the growth of SGC-7901 cells and induces cell apoptosis.