目的探讨环杷明(Cyclopamine)对胰腺癌细胞系ASPC-1增殖和凋亡的作用机制。方法不同浓度环杷明处理胰腺癌细胞系ASPC-1后,通过噻唑蓝(MTT)比色法试验检测癌细胞增殖状况,并计算不同浓度和作用时间的抑制率;用流式细胞仪检测各组的细胞周期,计算增殖指数(PI)和凋亡指数(AI);用裸鼠移植瘤模型验证环杷明的体内抑制作用。结果随着环杷明作用浓度增加和时间延长,对ASPC-1细胞增殖的抑制作用逐渐增强,当环杷明浓度达到10mol/L时,24h抑制率达到近90%。环杷明使细胞周期阻滞在G0/G1期,细胞凋亡率增加。环杷明浓度为10mol/L时,AJ达到25%左右,而PI下降至24%左右。在裸鼠移植瘤模型中,环杷明同期给药组肿瘤生长缓慢,而延期给药组开始生长速度与对照组相似,给药后生长逐渐受抑制,3组肿瘤均值间差异均有统计学有意义(P〈0.05)。结论环杷明通过细胞周期阻滞和促进细胞凋亡,发挥对胰腺癌细胞增殖的抑制效应,并且这种效应呈剂量和时间依赖性。
Objective To investigate the inhibition and apoptosis of pancreatic cancer cell line ASPC-1 induced by cyclopamine. Methods The ASPC-1 cells were treated with different concentrations of cyclopamine for different duration. The MTT assay was used to examine the antiproliferative effect of cyclopamine, and the inhibition rate was calculated. Flow cytometry (FCM) was used to examine the cell cycle of every group,and the proliferation index (PI) and apoptosis index (AI) were calculated. The inhibitory effect of cyclopamine in nude mice model was validated. Results The inhibition rate of ASPC-1 cells prolieration by cyclopamine was increased in a concentration- and time-dependent manner. When the concentration of cyclopamine reached 10 mol/L, the 24 h inhibition rate reached nearly 90%. Cyclopamine caused cell cycle arrest at G0/G1 phase and the increase of apoptosis. When the concentration of cyclopamine was 10 mol/L,the PI was decreased to 24% and AI increased to 25%. In the nude mice model, cyclopamine produced significant antiproliferative effect in concurrent group, but had weaker effect in delayed group (P 〈 0.05 ). Conclusion Cyclopamine could induce apoptosis and block cell cycle, and pro- duce time- and concentration-dependent antiproliferative effects in ASPC-1 cells.