灵芝总皂苷对结肠癌HCT116细胞的抑制作用

Inhibition of Colon Cancer Cell Line HCT116 by Total Saponins from Ganoderma lucidum

为探究灵芝总皂苷提取物对结肠癌HCT116细胞的抑制作用和相关机制,以灵芝子实体为原料,醇提并纯化总皂苷,通过CCK8法测定灵芝总皂苷提取物对结肠癌HCT116细胞的抑制效果,同时采用流式细胞术检测细胞凋亡,荧光分光光度计检测细胞内线粒体膜电位及细胞内活性氧含量,实时荧光定量PCR检测凋亡相关基因p53、noax、bax、bcl-2、caspase-9和caspase-3的表达。结果显示,与空白组相比,灵芝总皂苷质量浓度为40 mg/L以上时,可极显著地抑制HCT116细胞增殖,且抑制效果与时间呈正相关。经灵芝总皂苷干预后,细胞凋亡率显著增加,当加入灵芝总皂苷的质量浓度为150 mg/L时,其凋亡率达到50.2%,且灵芝总皂苷可显著降低细胞内线粒体膜电位,增加细胞内活性氧含量,增加促凋亡基因p53、noax、bax、caspase-9和caspase-3的表达,降低抗凋亡基因bcl-2的表达。综上,灵芝总皂苷对于结肠癌细胞具有显著抑制作用,且其抑制效果可通过线粒体凋亡途径实现,是值得进一步研究的抑结肠癌候选药物和食品添加剂。

This study aimed to explore the inhibitory effect and related mechanisms of the extracted Lucid Ganoderma total saponins(LGTS)on colon cancer HCT116 cells.Ganoderma lucidum fruiting bodies were used as the raw materials,and the LGTS was extracted by alcohol and then purified.The inhibitory effect of LGTS on colon cancer HCT116 cells was detected by CCK8 method.Meanwhile,the state of cell apoptosis was detected by flow cytometry.The intracellular mitochondrial membrane potential and intracellular reactive oxygen species levels were recorded by fluorescence spectrophotometer.The expression of apoptosis-related genes such as p53,noax,bax,bcl-2,caspase-9 and caspase-3 were measured by real-time fluorescent quantitative PCR.The results showed that the concentration of LGTS above 40 mg/L could significantly inhibit the proliferation of HCT116 cells compared with the blank group,with the inhibitory effect becoming positively correlated with time.Through the intervention of GLTS,the apoptosis rate of cells increased significantly.At a GLTS concentration of 150 mg/L,the apoptosis rate reached 50.2%.Additionally,GLTS significantly reduced the intracellular mitochondrial membrane potential,increased the intracellular reactive oxygen species levels,upregulated the expression of pro-apoptotic genes(p53,noax,bax,caspase-9 and caspase-3),and downregulated the expression of anti-apoptotic gene(bcl-2).In conclusion,GLTS have the significant inhibitory effect on colon cancer cells and the effect could be achieved through the mitochondrial apoptosis pathway,suggesting GLTS as promising candidates for further research as potential anti-colon cancer drugs and food additives.