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目的:观察扬子铁线莲中提取的三萜类皂苷成分对人胃癌SGC-7901细胞株生长及凋亡的作用,研究其对核转录因子NF-κB活性的影响,初步探讨其可能的作用机制。方法:应用不同浓度铁线莲皂苷作用于SGC-7901细胞48h后,MTT法检测细胞生长抑制率;流式细胞仪Annexin V/PI双染法检测联合应用铁线莲皂苷及5-氟尿嘧啶(5-FU)后SGC-7901细胞凋亡率;应用Western blot方法检测各组胞核中NF-κBp65表达情况;酶联免疫吸附试验(ELISA)进一步确定胞核内NF-κB的活性。结果:铁线莲皂苷各浓度组对胃癌SGC-7901细胞生长抑制率均明显高于对照组(P<0.05),且随药物浓度的增加而上升。铁线莲皂苷对5-FU诱导的细胞凋亡有明显的促进作用。Western blot及ELISA法均证实铁线莲皂苷对胞核NF-κB表达及活性有一定的抑制作用,并且这种抑制作用存在着剂量依赖现象。结论:铁线莲皂苷对胃癌SGC-7901细胞具有抑制增殖、诱导凋亡的作用,其诱导细胞凋亡的机制可能与抑制胞核NF-κB表达有关。
OBJECTIVE: To observe the effect of triterpene saponin extract on the growth and apoptosis of human gastric cancer cell line SGC-7901, and to study its effect on the activity of nuclear factor-κB . METHODS: SGC-7901 cells were treated with different concentrations of clematis for 48 hours. MTT assay was used to detect the cell growth inhibition rate. Flow cytometry Annexin V / PI double staining was used to detect clematis and 5-fluorouracil -FU). The protein expression of NF-κBp65 in nucleus of each group was detected by Western blot. The activity of NF-κB in nucleus was further determined by enzyme linked immunosorbent assay (ELISA). Results: The inhibitory rates of clematis saponin in gastric cancer SGC-7901 cells were significantly higher than those in control group (P <0.05), and increased with the increase of drug concentration. Clematis saponin could obviously promote 5-FU-induced apoptosis. Both Western blot and ELISA confirmed that Clematis saponin could inhibit the expression and activity of nuclear NF-κB in a dose-dependent manner. The inhibitory effect was dose-dependent. Conclusion: Clematis sativus can inhibit the proliferation and induce apoptosis of gastric cancer cell line SGC-7901, and its mechanism of inducing apoptosis may be related to the inhibition of nuclear NF-κB expression.