四种重楼皂苷对结肠癌细胞的体外毒性作用比较

摘要

目的：探讨重楼皂苷Ⅰ、Ⅴ、Ⅵ、H对结肠癌细胞的体外毒性，为寻求有效的结肠癌化疗药物提供依据。方法选取复苏结肠癌细胞系SW620、SW480、Colo320、Lovo，将其与重楼皂苷Ⅰ、Ⅴ、Ⅵ、H、奥沙利铂等药物共培养48 h，加CCK-8孵育2 h，酶标仪检测450 nm处吸光度值。采用SPSS23．0统计软件分析重楼皂苷对结肠癌细胞的毒性。 Annexin V／PI染色，流式细胞仪检测凋亡细胞。 Western blotting法检测Bax、Caspase-3蛋白表达。结果重楼皂苷Ⅰ、Ⅴ、Ⅵ、H对结肠癌细胞系SW620、SW480、Colo320、Lovo抗肿瘤活性均良好。四种重楼皂苷单体对结肠癌细胞IC50值波动于0．4～8 mg／L。重楼皂苷I和重楼皂苷V对结肠癌细胞的毒性强于另外两种重楼皂苷单体（P＜0．05）。重楼皂苷I（IC50值波动在0．4～0．6 mg／L）与奥沙利铂（IC50值0．1～2．1 mg／L）体外抗肿瘤作用相当。流式细胞仪显示重楼皂苷I处理的细胞Annexin V／PI阳性细胞比例升高52．3％，Western blotting结果显示重楼皂苷I处理的细胞Bax和Caspase-3蛋白表达分别是对照组的3倍和1．13倍。结论重楼皂苷Ⅰ、Ⅴ、Ⅵ、H对结肠癌细胞均有良好的抗肿瘤活性，其中以重楼皂苷I作用最强，其能介导结肠癌细胞凋亡。%Objective To investigate the in vitro toxicity of polyphyllins I, V, VI and H on colon cancer cells and to provide guidance for seeking effective chemotherapy drugs of colon cancer.Methods The resurgent SW620, SW480, Co-lo320, Lovo colon adenocarcinoma cell lines were co-cultured with polyphyllin I, polyphyllin VI, polyphyllin H, polyphyl-lin V and oxaliplatin for 48 h respectively, which were then added with CCK-8 for two-hour incubation.The optical density at 450 nm was detected by microplate reader.The cytotoxicity of polyphyllin on colon cancer cells was analyzed by SPSS 23.0.Annexin V/PI staining and flow cytometry was used to detect the apoptotic cells.Western blotting was applied to de-tect Bax and Caspase-3 protein expression.Results The polyphyllin I, polyphyllin VI, polyphyllin H and polyphyllin V showed remarkable antineoplastic activity on SW620, SW480, Colo320 and Lovo cell lines with IC50 of 0.4 mg/L to 8 mg/L.The anticancer activity of polyphyllin I and polyphyllin V were stronger than that of polyphyllin VI and polyphyllin H ( P<0.05).The in vitro anti-tumor effect of polyphyllin I with IC50 of 0.4-0.6 mg/L was the same as oxaliplatin with IC50 of 0.1-2.1 mg/L.Flow cytometry demonstrated that the percentage of Annexin V/PI positive cells treated with polyphyllin I increased 52.3%.Western blotting showing that polyphyllin I mediated expression of Bax and Caspase-3 in colorectal cells was 3 and 1.13 times higher than that of the control group.Conclusion Polyphyllin I, polyphyllin VI, polyphyllin H and polyphyllin V show good anticancer activity against colon adenocarcinoma cell lines in vitro, especially polyphyllin I which can mediate the apoptosis of colon cancer cells.