马钱子碱通过调控线粒体凋亡途径诱导人胰腺癌CFPAC-1细胞凋亡  

Brucine in inducing cell apoptosis of human pancreatic cancer CFPAC-1 cells by regulating the pathway of mitochondrial apoptosis

作　　者：田龙夫 张琦 崔立华 杨磊 周易[1] 田余 马波[3] Tian Longfu;Zhang Qi;Cui Lihua;Yang Lei;Zhou Yi;Tian Yu;Ma Bo(Department of Oncology,Huanggang Central Hospital,Huanggang 438000,China;Tianjin Key Laboratory of Organ Injury in Acute Abdominal Injury,Tianjin Hospital o f 1TCWM Nanhai Hospital,Tianjin 300100,China;Department of Oncology SurgeryⅡ,Tianjin Hospital of ITCWM Nankai Hospital,Tianjin 300100,China)

机构地区：[1]黄冈市中心医院肿瘤科,黄冈438000 [2]天津市中西结合医院南开医院天津市急腹症器官损伤中西医结合修复重点实验室,天津300100 [3]天津市南开医院肿瘤外Ⅱ科,天津300100

出　　处：《中华胰腺病杂志》2019年第6期425-429,共5页Chinese Journal of Pancreatology

基　　金：国家自然科学基金(81602496)。

摘　　要：目的探讨马钱子碱对人胰腺癌CFPAC-1细胞凋亡的影响及其可能机制。方法以不同浓度马钱子碱干预CFPAC-1细胞。采用MTT比色法和流式细胞仪检测细胞增殖和凋亡;采用花青染料(JC-1)染色法检测细胞线粒体膜电位的变化;采用蛋白质印迹法检测Bax、B cl-2蛋白表达情况。结果〇(对照组)及〇.4、0.8 m m ol/L马钱子碱组干预CFPAC-1细胞24、48、72 h后,细胞增殖抑制率分别为0;(30.23±0.55)%、(40.61±0.15)%、(46.98±1.27)%;(50.17±0.75)%、(61.23±0.91)%、(70.32±0.40)%,呈浓度及时间依赖性增加而升高,显著高于对照组,且不同时间两两组间比较差异均有统计学意义(/>值均<〇.〇5)。〇及0.4、0.8 mmol/L马钱子碱干预CFPAC-1细胞48 h后,细胞凋亡率分别为(2.92±0.46)%、(4.64±1.31)%、(13.09±0.65)%,随药物浓度增加而逐渐升高,其中0.8 mm ol/L干预组显著高于对照组,差异有统计学意义(P<0.05);随着药物浓度的增加,呈红色荧光的未凋亡活细胞逐渐减少,而呈绿色荧光的凋亡坏死细胞逐渐增多,提示CFPAC-1细胞线粒体膜电位受到重度破坏而降低;CFPAC-1细胞的B d-2蛋白表达量分别为(0.92±0.12)、(0.67±0.14)、(0.35±0.14)mmol/L,Bax蛋白表达量分别为(0.56±0.12)、(0•85±0.10)、(1.15±0.12)m m ol/L,随马钱子碱浓度增加,Bcl-2表达量显著下调,而Bax表达量显著上调,差异均有统计学意义(P值均<0.05)。结论马钱子碱可能通过线粒体凋亡途径上调Bax和下调Bcl-2表达,从而诱导人胰腺癌CFPAC-1细胞凋亡。Objective To investigate the influence of Brucine on cell apoptosis of pancreatic cancer CFPAC-1 cells and the possible mechanism.Methods Brucine in different concentrations were used to treat CFPAC-1 cells.Cell proliferation was determined by MTT assay and cell apoptosis was determined by flow cytometer assay.Mitochondrial membrane potential was examined by JC-1 staining.The protein expression of Bax and Bcl-2 was measured by Western Blot.Results The growth inhibition rates of CFPAC-1 cells after being treated with 0(control group),0.4 and 0.8 mmol/L Brucine for 24,48 and 72 h were 0,(30.23±0.55)%,(40.61±0.15)%,(46.98±1.27)%a n d(50.17±0.75)%,(61.23±0.91)%,(70.32±0.40)%,increasing with a concentration-and time-dependent increase,which was higher than that in control group;and the differences between either two groups at different time points were statistically significant(P<0.05).CFPAC-1 cell apoptosis rate after being treated with 0,0.4 and 0.8 mmol/L Brucine for 48 h was(2.92±0.46)%,(4.64±1.31)%and(13.09±0•65)%,which increased gradually with the increased drug concentration.The apoptotic rate in 0.8 mmol/L treatment group was obviously higher than that in control group,and the difference was statistically significant(P<0.05).With the increase of the drug concentration,the red fluorescence gradually decreased,and the green fluorescence gradually increased,indicating that the mitochondrial membrane potential was severely damaged and thus decreased.The protein expression of Bcl-2 in CFPAC-1 cells were(0.92±0.12),(0.67±0.14)and(0.35±0.14)mmol/L,and the expression of Bax in CFPAC-1 cells were(0.56±0.12),(0.85±0.10)and(1.15±0.12)mmol/L.With the increase of brucine concentration,the expression of Bcl-2 was significantly reduced while the expression of Bax was significantly increased;and the difference was statistically significant(P<0.05).Conclusions Brucine can effectively induce the apoptosis of human pancreatic cancer CFPAC-1 cells through mitochondrial apoptotic pathway by up-regulating the expre

关 键 词：胰腺肿瘤 马钱子碱 CFPAC-1细胞 细胞凋亡 线粒体途径 

分 类 号：R28[医药卫生—中药学]