蟾毒灵介导JNK信号通路诱导人胰腺癌细胞的凋亡  

Bufalin induces apoptosis of pancreatic cancer cells via the JNK signaling pathway

作　　者：胡强[1] 殷佩浩[2] 陆品相[1] 沈玉根[1] 余琛[3] 李水军[3] 贾晶莹[3] 刘罡一[3] 缪怡[3] 范耀祖[4] 

机构地区：[1]上海市徐汇区中心医院普外科,上海市200031 [2]上海市普陀区中心医院普外科,上海市200062 [3]上海市徐汇区中心医院中心实验室,上海市200031 [4]上海市同济医院普外科,上海市200442

出　　处：《世界华人消化杂志》2012年第2期100-105,共6页World Chinese Journal of Digestology

基　　金：江苏大学课题基金资助项目;No.JLY20080093~~

摘　　要：目的:研究蟾毒灵诱导人胰腺癌细胞凋亡以及凋亡相关基因表达的JNK信号转导通路,揭示其抗胰腺癌的部分机制.方法:MTT法观察蟾毒灵对人胰腺癌BxPC-3细胞的生长抑制作用;0.16、0.32、0.64mg/L蟾毒灵分别作用人胰腺癌BxPC-3细胞48h后,流式细胞仪(flow cytometry,FCM)检测细胞周期和细胞凋亡;Western印迹法检测蟾毒灵作用BxPC-3细胞后SAPK/JNK信号通路的激活情况,荧光定量PCR检测Survivin基因mRNA的表达水平;并比较阻断JNK信号通路后丹参酮ⅡA对胰腺癌细胞凋亡Survivin基因mRNA的表达.结果:MTT法测得蟾毒灵对人胰腺癌BxPC-3细胞具有显著的抑制作用,其作用效果与剂量和作用时间成正相关;0.16、0.32、0.64mg/L浓度蟾毒灵作用人胰腺癌细胞后的细胞凋亡率分别为19.36%±0.39%、40.69%±0.44%、59.63%±1.14%,与对照组2.24%±0.37%比较均有显著性差异(P<0.01);蟾毒灵作用人胰腺癌细胞1h后JNK信号通路被激活,2h达峰值;阻断JNK信号通路后,凋亡率明显降低(P<0.01);0.32mg/L蟾毒灵作用人胰腺癌细胞48h后Survivin mRNA的表达明显下降;阻断JNK信号通路后,蟾毒灵作用人胰腺癌细胞的Survivin mRNA的表达明显上升.结论:蟾毒灵通过JNK信号转导通路下调人胰腺癌BxPC-3细胞Survivin mRNA的表达,可能是其诱导胰腺癌细胞凋亡的机制.AIM：To investigate whether bufalin induces apoptosis of human pancreatic cancer cells via the JNK signaling pathway.METHODS：The cytostatic effect of bufalin on human pancreatic cancer cell line BxPC-3 was evaluated by MTT assay.The impact of bufalin on apoptosis and p-JNK expression was determined by flow cytometry （FCM） and Western blot,respectively.The mRNA expression level of survivin was detected by fluorescent quantitative PCR in BxPC-3 cells treated with bufalin in the absence or presence of a JNK inhibitor.RESULTS：Bufalin inhibited cell growth in a concentrationand time-dependent manner.BxPC-3 cells treated with 0.16,0.32,or 0.64mg/L of bufalin for 48 h had cell apoptotic rates of 19.36%±0.39%,40.69%±0.44% and 59.63%±1.14%,respectively,showing a significant difference among the three groups （P0.01）.After the JNK signaling pathway was blocked,cell apoptotic rate decreased significantly （P0.01）.After bufalin treatment,p-JNK expression was upregulated at 1 h and reached the peak at 2 h.The mRNA expression of survivin decreased obviously after treatment with 0.32mg/L of bufalin for 48 h,but increased significantly when the NK signaling pathway was blocked.CONCLUSION：Bufalin can induce apoptosis of human pancreatic cancer cells.The anti-pancreatic cancer activity of bufalin might involve down-regulation of survivin mRNA via the SAPK/JNK signal transduction pathway.

关 键 词：蟾毒灵 胰腺癌 凋亡 信号转导 

分 类 号：R735.9[医药卫生—肿瘤]