四种重楼皂苷对结肠癌细胞的体外毒性作用比较

In vitro toxicity of four kinds of polyphyllins on colon cancer cells

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,Colo320,Lovo colon adenocarcinoma cell lines were co-cultured with polyphyllin I,polyphyllin VI,polyphyllin H,polyphyllin 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 SPSS23. 0. Annexin V / PI staining and flow cytometry was used to detect the apoptotic cells. Western blotting was applied to detect 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 IC50of0. 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.