白头翁皂苷抑制人HT29结肠癌细胞增殖及诱导凋亡作用研究

Effects of Pulsatilla chinensis saponins on proliferation and apoptosis of HT29 cells

目的:考察白头翁皂苷对人结肠癌细胞株HT29的增殖及诱导细胞凋亡的作用。方法:MTT法检测白头翁皂苷对HT29细胞增殖的影响;DAPI染色、Annexin-V/PI染色流式细胞术检测对细胞凋亡的影响;Western blotting检测白头翁皂苷对凋亡相关蛋白cleaved-caspase3的影响。结果:白头翁皂苷能明显抑制HT29细胞的增殖,其20、10、5、2.5、1.25、0.625μg/ml浓度作用48h对HT29增殖抑制率依次为97.6%、87.1%、76.0%、67.3%、59.4%、41.9%,呈浓度依赖关系,IC50值为1.440μg/ml;DAPI染色可以观察到白头翁皂苷干预后的HT29细胞出现明显的凋亡形态;流式细胞术检测白头翁皂苷5、10、15、20μg/ml诱导HT29细胞凋亡率依次为10.13%、10.93%、13.17%、13.63%;Western blotting检测白头翁皂苷药物(2.5、5、10、15、20μg/ml)干预后HT29细胞出现Cleaved-caspase 3的表达,呈浓度依赖关系,在20μg/ml与对照组比较有显著性差异。结论:白头翁皂苷可明显抑制HT29细胞增殖,其抗肿瘤作用与诱导肿瘤细胞凋亡有关。

This study was to investigate the effect of Pulsatilla saponins on proliferation and apoptosis of HT29 cells. Methods： HT29 cells were treated with Pulsatilla saponins,cell viability was assessed by MTT assay; apoptosis were observed by DAPI staining,the apoptosis ratio were assessed by flow cytometry after Annexin V/PI staining;Cleaved-caspase3 were detected by Western Blotting assay. Result： Pul- satilla saponins could inhibit the proliferation of HT29 cells in a manner of dosedepedence, the inhibition rate of proliferation at the dose of 20,10,5,2.5,1.25,0.625 μg/ml is 97.6%, 8;7.1% ,76.0%, 67.3%, 59.4% ,41.9% ; Apoptotic morphology could be observed by DAPI staining; the apoptosis ratio of HT29 cells treated with Pulsatilla saponins 48hr at the dose 5,10,15,20μg/ml was 10. 13%, 10. 93%, 13. 17% , 13.63% ; PulsatiUa saponins（ 2.5,5,10,15,20μg/ml） could increase the expression of cleaved easpase3 in HT29 in a manner of dose depedence, and it showed a significant difference compared with control group at the dose of 20μg/ml. Conclusion： Pulsatilla saponins could inhibit the proliferation of HT29 cells, show a strong anti-tumor effect in vitro,the mechanism was related with apoptosis.