龙胆苦苷对人结肠癌HCT116细胞增殖、凋亡的影响

Effect of gentiopicroside on human colorectal cancer HCT116 cell proliferation and apoptosis

目的研究龙胆苦苷对人结肠癌细胞HCT116增殖、凋亡的影响,并初步探讨其可能作用机制。方法体外培养人结肠癌HCT116细胞,采用不同浓度的龙胆苦苷(0、12.5、25、50 mg/L)作用于细胞,并同时应用5-氟尿嘧啶作用于相同的细胞,对比不同处理方法对细胞生长的抑制及对细胞形态及凋亡的影响,并检测不同浓度龙胆苦苷对细胞凋亡相关基因Caspase-3、Bax、Bcl-2表达水平的影响。结果与阴性对照组相比,5-氟尿嘧啶组和龙胆苦苷组均显著能够抑制HCT116细胞增殖,且龙胆苦苷组对HCT116细胞的抑制增殖作用呈剂量和时间依赖性,差异有统计学意义(P<0.05)。5-FU组凋亡细胞百分率最高,其次是龙胆苦苷50、25、12.5 mg/L组,与阴性对照组比较,龙胆苦苷组(50、25、12.5 mg/L)细胞凋亡率显著提高,差异有统计学意义(P<0.05)。与阴性对照组比较,龙胆苦苷(12.5、25、50 mg/L)组Bax、Caspase-3 mRNA表达水平显著升高,而Bcl-2 mRNA表达在25、50 mg/L龙胆苦苷处理后明显降低,差异均有统计学意义(P<0.05)。与阴性对照组比较,龙胆苦苷(12.5、25、50 mg/L)组促凋亡蛋白Bax、Caspase-3的表达水平明显上调,而抑凋亡蛋白Bcl-2的表达明显下调,差异均有统计学意义(P<0.05)。结论龙胆苦苷可抑制HCT116细胞增殖并促进细胞凋亡,且对肿瘤细胞的抑制增殖作用呈剂量和时间依赖性,可能与激活Bax表达、抑制Bcl-2表达进而活化Caspase-3信号通路有关。

Objective To investigate the effect of gentiopicroside on the proliferation and apoptosis of human colorectal cancer cell HCT116,and explore the potential mechanism.Methods HCT116 cells were cultured in vitro,and treated with respectively gentiopicroside at different concenntrations.At the same time,5-fluorouracil was used to treat the same cells,and the inhibition of cell growth and the influence of different treatment methods on cell morphology and apoptosis were compared,and the effects of different concentrations of gentiopicrin on the expression levels of apoptosis related genes Caspase-3,Bax,Bcl-2 were detected.Results Compared with the control group,both 5-fluorouracil group and gentiopicrin group could significantly inhibit the proliferation of HCT116 cells,and gentiopicrin group had a dose-dependent and time-dependent inhibitory effect on the proliferation of HCT116 cells.The 5-FU group had the highest percentage of apoptotic cells,followed by the gentiopicroside 50,25,and 12.5 mg/L groups,and the difference was statistically significant(P<0.05).Compared with the negative control group,the gentiopicroside group(50,25,and 12.5 mg/L)had the highest percentage of apoptosis The rate was significantly improved,and the difference was statistically significant(P<0.05).Compared with the negative control group,the expression levels of Bax and Caspase-3 mRNA in the gentiopicroside(12.5,25,and 50 mg/L)groups were significantly increased,while the expression levels of Bcl-2 mRNA were significantly increased in the gentiopicroside(12.5,25,and 50 mg/L)groups.After treatment,it decreased significantly,and the difference was statistically significant(P<0.05).Compared with the negative control group,the expression levels of pro-apoptotic proteins Bax and Caspase-3 in the gentiopicroside(12.5,25,and 50 mg/L)group were significantly up-regulated,while the expression of anti-apoptotic protein Bcl-2 was significantly down-regulated,and the difference was statistically significant(P<0.05).Conclusion Gentiopicroside could inhibit the proliferation of human colorectal cancer cell HCT116 cells and induce cell apoptosis,which may be related to regulating Bax and Bcl-2 expression and activating Caspase-3 signal pathway.