重楼皂苷Ⅰ联合加热抗胃癌MKN-45细胞株增殖的体外研究

Study on Antitumor Activity of Polyphyllin Ⅰ with Hyperthermia on Human Gastric Cancer Cell Lines MKN-45 in Vitro

目的:观察重楼皂苷Ⅰ(polyphyllin I,PPⅠ)联合加热抗胃癌MKN-45细胞株的效应。方法:PPⅠ与MKN-45细胞共培养,采用MTT法检测PPⅠ对细胞增殖的影响,流式细胞仪检测细胞凋亡变化;PPⅠ联合加热处理MNK-45细胞,集落形成试验检测加热对PPⅠ抗MNK-45细胞抗瘤活性的影响,Western blot法检测p21蛋白表达变化。结果:不同浓度PPⅠ作用于MKN-45胃癌细胞株24h和48h后,细胞存活率显著降低,呈时间和剂量依赖关系。PPⅠ作用于MKN-45细胞24 h后,MNK-45细胞凋亡率由(4.3±1.1)%升高至(37.8±3.5)%。PPⅠ(IC30)联合加热后,细胞存活率由(61.2±3.4)%降至(40.7±3.1)%(P<0.05)。PPⅠ能上调MKN45细胞p21蛋白表达,这一效应能被加热增强。结论:PPⅠ对胃癌MKN-45细胞株有明显增殖抑制和诱导凋亡的作用,可能与其上调p21蛋白表达有关,联合加热能增强这一效应。

Objective：To evaluate the antitumor activity of PPⅠ with hyperthermia on human gastric cancer cell lines MKN-45 in vitro.Methods：Human gastric cancer MKN-45 cells were cultured and treated with different concentrations of PPⅠ with or without hyperthermia.Cell proliferation and apoptosis were measured by MTT assay and flow cytometry.The influence of hyperthermia on the antitumor effects of PPⅠ was measured by colony formation assay.The p21 protein levels were assessed by western blot analyses.Results：The survival rates of MKN-45 cells were significantly decreased after 24 and 48 hours culturing with different concentrations of PPⅠ,and this effect exhibited time-dose-dependent.Apoptosis rate was increased from（4.3±1.1）% to（37.8±3.5）% after culturing with PPⅠ for 24 hours.Compared with PPⅠ alone,when combined with hyperthermia,cell survival rate was decreased from（61.2±3.4）% to（40.7±3.1）%（P〈0.05）.The expression of p21 protein can be upregulated by PPⅠ and this effect can be enhanced by hyperthermia.Conclusion：PPⅠ exhibits growth inhibition and apoptosis induction in human gastric cancer cell lines MKN-45,and the effects can be enhanced by hyperthermia.The antitumor mechanism may be related to upregulating the expression of p21 protein.