血根碱通过调控PI3K/Akt信号通路诱导胰腺癌细胞凋亡的机制  

Mechanism of Sanguinarine in Inducing Apoptosis in Murine Pancreatic Cancer Cells by Inhibiting PI3K/Akt Signaling Pathway

作　　者：田龙夫 张琦 王波涛 崔立华[2] 杨磊[2] 马波[2] 崔云峰[2] TIAN Long-fu;ZHANG Qi;WANG Bo-tao;CUI Li-hua;YANG Lei;MA Bo;CUI Yun-feng(Huanggang Central Hospital,Huanggang 438000,China;Tianjin Hospital of ITCWM Nankai Hospital,Tianjin 300100,China;Tianjin Medical University,Tianjin 300070,China)

机构地区：[1]黄冈市中心医院,湖北黄冈438000 [2]天津市南开医院,天津300100 [3]天津医科大学,天津300070

出　　处：《中国实验方剂学杂志》2018年第19期166-171,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81602496)

摘　　要：目的：探讨血根碱对胰腺癌细胞凋亡的影响及其信号通路调控机制。方法：不同浓度血根碱（2,4,6μmol·L-1）处理小鼠胰腺癌Panc02细胞不同时间（24,48,72 h）后,采用噻唑蓝（MTT）比色法检测血根碱对细胞生长抑制作用的影响;采用磷脂结合蛋白V/碘化丙啶（Annexin V/PI）双染色试剂盒通过流式细胞仪检测血根碱对Panc02细胞凋亡影响;采用花青染料（JC-1）探针试剂盒荧光染料分析检测线粒体膜电位的变化;采用蛋白免疫印迹法（Western blot）检测B淋巴细胞瘤-2（Bcl-2）,Bcl-2相关X蛋白（Bax）,总蛋白激酶B（Akt）,磷酸化蛋白激酶B（p-Akt）,总磷脂酰肌醇3激酶（PI3K）,磷酸化磷脂酰肌醇3激酶（p-PI3K）的表达情况。结果：与空白组比较,不同浓度的血根碱作用同一时间后,随着药物浓度的增加细胞生长抑制率逐渐升高（P〈0.05）,与空白组比较,同一浓度作用不同时间后,随着时间的增加细胞生长抑制率逐渐升高（P〈0.05）;与空白组比较,血根碱2μmol·L-1组的细胞凋亡率无明显升高,与空白组比较,血根碱4,6μmol·L-1组的细胞凋亡率均明显升高（P〈0.05）;与空白组比较,血根碱2μmol·L-1组时红绿荧光无变化,血根碱4,6μmol·L-1组红色荧光减弱,绿色荧光增强,Panc02细胞线粒体膜电位降低;与空白组比较,血根碱2μmol·L-1组处理的Bax,Bcl-2,p-PI3K,p-Akt,Akt和PI3K蛋白表达均无变化,血根碱6μmol·L-1组Bax蛋白表达增高,Bcl-2及PI3K/Akt信号通路中关键蛋白p-PI3K,p-Akt表达降低（P〈0.05）。结论：血根碱通过抑制PI3K/Akt信号通路有效地诱导小鼠胰腺癌Panc02细胞经线粒体凋亡途径发生凋亡。Objective： To investigate the effect of sanguinarine on pancreatic cancer cell apoptosis and its signal pathway regulation mechanism. Method： After Panc02 cells were treated with different concentrations of sanguinarine（ 2,4,6 μmol·L-1） for different periods of time（ 24,48,72 h）,the inhibition of cell growth was determined by 3-（ 4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2 H-tetrazolium bromide（ MTT） assay. The effect of sanguinarine on cell apoptosis was determined by phospholipid binding protein V/propidium iodide（ Annexin V-FITC/PI） double staining flow cytometer assay. The changes in mitochondrial membrane potential were examined by cyanine dye（ JC-1） staining assay. The expressions of protein B-lymphoblast-2（ Bcl-2）,Bcl-2 related X protein（ Bax）, total protein kinase B（ Akt）, phosphorylated protein kinase B（ p-Akt）, total phosphatidylinositol 3 kinase（ PI3 K）,phosphorylated phosphatidylinositol 3-kinase（ p-PI3 K） were measured by Western blot analysis. Result： Compared with the blank group,the growth inhibition rate of Panc02 cells increased gradually with the rise of the drug concentration,after being treated with different concentrations of sanguinarine for the same period of time（ P〈 0. 05）. Compared with the blank group,the inhibition rate of cell growth increased gradually with the rise of time,after being treated with the same concentration for different periods of time（ P〈 0. 05）. Compared with the blank group,the apoptosis rate was not obviously increased at the dose of the2 μmol·L-1 sanguinarine. Compared with the blank group,the apoptotic rate was increased at the dose of the 4,6μmol·L-1 sanguinarine（ P〈 0. 05）. Compared with the blank group,there was no change in the red and green fluorescence at the dose of 2 μmol·L-1,but a decrease in red fluorescence,an increase in green fluorescence and a reduction in Panc02 mitochondrial membrane potential at the dose of 4,6 μmol·L-1 sanguinarine. Compared with the blank group,the

关 键 词：胰腺癌 血根碱 细胞凋亡 Panc02细胞 磷脂酰肌醇3激酶/蛋白激酶B信号通路 

分 类 号：R735[医药卫生—肿瘤] R22[医药卫生—临床医学]