槐耳清膏对胰腺癌细胞增殖和迁移的影响

Effects of Huaier cream on proliferation and migration of pancreatic cancer cells

目的研究槐耳清膏在体外对人胰腺癌细胞增殖和迁移的影响及其机制。方法将对数生长期的PANC-1胰腺癌细胞用不同浓度的槐耳清膏溶液处理不同时间。应用荧光显微镜观察细胞的形态学变化,CCK-8法检测槐耳清膏对细胞增殖的影响,流式细胞术检测细胞凋亡及细胞周期的变化,划痕实验和Transwell实验观察细胞迁移和侵袭能力的变化,Western blot观察槐耳清膏对凋亡相关蛋白和上皮细胞-间充质转化相关蛋白表达的影响。结果槐耳清膏处理后的PANC-1细胞变大,形态不规则,细胞质内空泡增多,对细胞增殖的抑制率和凋亡率呈浓度和时间依赖性地增加(P<0.05)。细胞周期实验显示,槐耳清膏处理后的细胞阻滞于G1期,且其所占比例随药物浓度及作用时间的增加而增加(P<0.05)。槐耳清膏抑制细胞迁移,且呈浓度和时间依赖性(P<0.05)。随着槐耳清膏药物浓度的增加,细胞穿过基膜的数量减少(P<0.05)。随时间及槐耳清膏药物浓度的上升,Bad、Caspase-3、Caspase-9、磷酸化p53、上皮性钙黏附蛋白表达增强,Bcl-2、神经性钙黏附蛋白、Akt表达减弱(P<0.05)。结论槐耳清膏能抑制胰腺癌PANC-1细胞增殖,诱导细胞凋亡,减弱细胞侵袭和迁移能力,可能是通过磷脂酰肌醇3-激酶/Akt信号通路实现的。

Objective To study the effects of Huaier cream on the proliferation and migration of human pancreatic cancer cells in vitro and underlying mechanism.Methods PANC-1 pancreatic cancer cells in logarithmic growth phase were treated with different concentrations of Huaier cream for different times.Fluorescence microscopy was used to observe the morphological changes of the cells. CCK-8method was used to detect the effect of Huaier cream on cell proliferation.Flow cytometry was used to detect the apoptosis and cell cycle changes.Scratch test and Transwell test were used to observe cell migration and invasion.The effect of Huaier cream on the expressions of apoptosis-related proteins and epithelial-mesenchymal transition-related proteins was observed by Western blot.Results The PANC-1 cells treated with Huaier cream were enlarged,irregular in shape and increased in vacuoles in cytoplasm.The inhibition rate of cell proliferation and apoptotic rate increased in the concentration-and time-dependent manners (P<0.05).The cell cycle test showed that the cells treated by Huaier cream were arrested in G1 phase,and the proportion increased with the increase of drug concentration and time(P<0.05).Huaier cream inhibited cell migration in the concentrationand time-dependent manners (P<0.05).The number of cells passing through the basement membrane decreased with the increase of the concentration of Huaier cream (P<0.05).The expressions of Bad,Caspase-3,Caspase-9,phosphorylated p53and epithelial cadherin were enhanced, while the expressions of Bcl-2,neuronal cadherin and phosphorylated Akt were decreased with the increase of time and concentration of Huaier cream (P<0.05).Conclusion Huaier cream can inhibit the proliferation,induce apoptosis,and weaken invasion and migration ability of PANC-1 pancreatic cancer cells,which is probably through phosphatidylinositol 3-kinase/Akt signaling pathway.