灵芝提取物对4种人肿瘤细胞生长的抑制作用及机制研究

Study of anti-proliferation effect of ganoderma lucidum extracts on four human tumor cells and its mechanism

目的:探讨超临界二氧化碳结合乙醇提取的灵芝提取物(GLE)对肿瘤细胞增殖、细胞周期及凋亡的影响及分子机制。方法:CCK8法检测GLE对4种肿瘤细胞系A549、SW1990、SKOV3及HGC27增殖的影响;流式细胞术检测GLE对肿瘤细胞凋亡和细胞周期的影响;RT-PCR检测凋亡和细胞周期相关基因mRNA表达;Western blot检测相关蛋白表达。结果:GLE时间-剂量依赖性地抑制4种肿瘤细胞增殖,诱导肿瘤细胞凋亡,降低Bcl2/Bax比值,上调cleaved caspase-3表达及cleaved PARP/总PARP比值;GLE可诱导肿瘤细胞发生细胞周期阻滞,改变细胞周期相关基因E2F-1、CDK2、CDK4、CDK6、CyclinA2、CyclinB1、CyclinE1及p21 mRNA表达。结论:GLE可显著抑制肿瘤细胞增殖,诱导细胞凋亡和细胞周期阻滞,其机制可能为细胞周期相关基因水平改变及caspase-3信号通路改变。

Objective:To investigate effect of ganoderma lucidum extracts(GLE),which was extracted by supercritical carbon dioxide combined with ethanol,on tumor cell proliferation,cell cycle and apoptosis and its molecular mechanisms.Methods:CCK8 assay was used to detect effect of GLE on proliferation of A549,SW1990,SKOV3 and HGC27 tumor cells.Flow cytometry was used to detect effect of GLE on tumor cell apoptosis and cell cycle;mRNA expressions of apoptosis and cell cycle related genes were detected by RT-PCR.Expressions of related proteins were detected by Western blot.Results:GLE inhibited proliferation of four tumor cells in a time and dose-dependent manner,induced apoptosis of tumor cells,decreased Bcl-2/Bax ratio,while increased cleaved caspase-3 expression and cleaved PARP/total PARP ratio;GLE can induce cell cycle arrest of tumor cells and change mRNA expressions of cell cycle related genes including E2F-1,CDK2,CDK4,CDK6,CyclinA2,CyclinB1,CyclinE1 and p21.Conclusion:GLE can significantly inhibit proliferation of tumor cells,induce apoptosis and cell cycle arrest,whose mechanism may be related to changes of expressions of cell cycle related genes and caspase-3 signaling pathway.