重楼皂苷Ⅶ对胰腺癌细胞的作用研究  

The Effect of Polyphyllin Ⅶ on Pancreatic Carcinoma Cells

作　　者：胡世尚 周圆圆 吴子豪 朱敏[1] 郗雪艳[1] 杜伯雨[1] 喻青青 HU Shi-shang;ZHOU Yuan-yuan;WU Zi-hao;ZHU Min;XI Xue-yan;DU Bo-yu;YU Qing-qing(Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院,湖北十堰442000

出　　处：《湖北医药学院学报》2020年第6期544-550,F0002,共8页Journal of Hubei University of Medicine

基　　金：十堰市科技局项目-引导性项目(19Y16);湖北省大学生创新创业项目(S201913249008);湖北医药学院2018年研究生启动金(2018QDJR31)。

摘　　要：目的:研究重楼皂苷Ⅶ对胰腺癌Miapaca-2细胞增殖、迁移及凋亡的影响。方法:不同浓度的重楼皂苷Ⅶ处理胰腺癌细胞后,高内涵筛选(High-content screening,HCS)分析细胞数量、面积、迁移速度、圆度的变化;MTT法检测细胞存活能力;平板克隆技术检测细胞增殖能力;划痕实验和Transwell观察重楼皂苷Ⅶ抑制细胞的迁移能力;DAPI染色和流式细胞术检测细胞形态和凋亡情况;成球实验检测重楼皂苷Ⅶ对Miapaca-2干细胞的抑制情况;Real-time PCR检测重楼皂苷Ⅶ处理Miapaca-2干细胞标志物CD133的表达情况。结果:MTT结果显示重楼皂苷Ⅶ能明显抑制胰腺癌Miapaca-2细胞的活力,呈剂量依赖关系;重楼皂苷Ⅶ对胰腺癌细胞迁移、侵袭有一定的抑制作用;DAPI染色发现随着药物浓度升高,细胞形态发生明显变化,出现核固缩;流式结果表明细胞凋亡率随重楼皂苷Ⅶ浓度增大而逐渐升高;重楼皂苷Ⅶ可抑制Miapaca-2的成球能力;同时可导致Miapaca-2干细胞标志物CD133的表达降低。结论:重楼皂苷Ⅶ在体外可以抑制胰腺癌Miapaca-2细胞的增殖、迁移和侵袭能力,诱导细胞发生凋亡,并且抑制胰腺癌干细胞的增殖及干细胞标志物CD133的表达。Objective To investigate the effects of polyphyllinⅦon the proliferation,migration and apoptosis of pancreatic carcinoma Miapaca-2 cells.Methods After the pancreatic carcinoma Miapaca-2 cells were treated with different concentrations of polyphyllin Ⅶ,high content screening(HCS)was used to analyze the changes of cell number,area,migration speed and roundness.MTT method was used to detect cell viability,and plate cloning technology was used to detect cell proliferation.Scratch test and Transwell were used to detect the inhibitory effect of PolyphyllinⅦon the migration of Miapaca-2 cells.The cell morphology and apoptosis were detected by DAPI staining and flow cytometry.Spheroidization test was used to detect the inhibition of Polyphyllin Ⅶ on Miapaca-2 stem cells,and Real-time PCR was used to detect the expression of stem cell marker CD133 in Miapaca-2 cells treated with Polyphyllin Ⅶ.Results MTT results showed that polyphyllinⅦcould significantly inhibit the activity of Miapaca-2 cells in a dose-dependent manner;Polyphyllin Ⅶ can inhibit the migration and invasion of Miapaca-2 cells;DAPI staining showed that,with the increase of drug concentration,cell morphology changed significantly,and karyopyknosis appeared;and flow cytometry showed that the apoptosis rate increased with the increase of polyphyllinⅦconcentration;polyphyllin Ⅶ could inhibit the ability of Miapaca-2 spheroidizaation,and reduce the expression of miapaca-2 stem cell marker CD133.Conclusion Polyphyllin Ⅶ can inhibit the proliferation,migration and invasion of pancreatic carcinoma Miapaca-2 cells in vitro,induce apoptosis,and inhibit the proliferation of pancreatic carcinoma stem cells and the expression of stem marker CD133.

关 键 词：重楼皂苷Ⅶ 胰腺癌 MIAPACA-2 细胞增殖 细胞凋亡 肿瘤干细胞 

分 类 号：R285.5[医药卫生—中药学]