广藿香酮抑制人胆囊癌SGC-996细胞的增殖  

Patchoulenone inhibits the proliferation of human gallbladder carcinoma SGC-996 cells

作　　者：吴瑶诗 黄缘[2] 董家鸿[2] 

机构地区：[1]南昌大学第二附属医院消化内科,江西南昌330006 [2]清华大学附属北京清华长庚医院肝胆胰中心,北京102218

出　　处：《肿瘤》2017年第1期50-57,共8页Tumor

摘　　要：目的 :探讨广藿香酮对人胆囊癌SGC-996细胞增殖和凋亡的影响及其可能的作用机制。方法:MTT法检测10、40、80、100和120μg/m L广藿香酮作用24、48和72 h后,人胆囊癌SGC-996细胞的增殖情况。克隆形成实验检测10、20和30μg/m L广藿香酮对胆囊癌SGC-996细胞克隆形成的影响。FCM法检测40、60和80μg/m L广藿香酮作用48 h后,胆囊癌SGC-996细胞的凋亡及细胞周期分布。蛋白质印迹法检测40、60和80μg/m L广藿香酮作用胆囊癌SGC-996细胞48 h后,活化型caspase-9、活化型caspase-3、剪切型多聚ADP核糖聚合酶1[ploy(ADP-ribose)ployrnerase-1,PARP-1]、Bax、Bcl-2及cyclin D1、cyclin A、cyclin B1蛋白的表达水平。结果:10、40、80、100和120μg/m L广藿香酮可明显抑制人胆囊癌SGC-996细胞的增殖(P值均<0.01),且呈剂量和时间依赖性(P值均<0.05)。10、20和30μg/m L广藿香酮能明显抑制SGC-996细胞的克隆形成(P值均<0.05)。40、60和80μg/m L广藿香酮可显著促进SGC-996细胞的凋亡(P值均<0.05),并将细胞周期阻滞于S期(P值均<0.05)。40、60和80μg/m L广藿香酮处理后,SGC-996细胞中活化型caspase-9、活化型caspase-3、剪切型PARP-1蛋白表达水平及Bax/Bcl-2值均显著上调(P值均<0.05),而cyclin D1、cyclin A和cyclin B1蛋白的表达水平则显著下调(P值均<0.01)。结论 :广藿香酮可抑制胆囊癌SGC-996细胞的增殖,并将细胞周期阻滞于S期。这一作用可能与广藿香酮调控线粒体凋亡途径相关蛋白及细胞周期相关蛋白的表达水平有关。Objective： To investigate the effects of patchoulenone on the proliferation and apoptosis of human gallbladder carcinoma cell line SGC-996, and to explore its possible mechanism.Methods： The proliferation of gallbladder carcinoma SGC-996 cells after treatment with 10, 40, 80, 100 and 120 μg/mL patchoulenone for 24, 48 and 72 h was detected by MTT assay. The effect of 10, 20 and 30μg/mL patchoulenone on the colony formation of human gallbladder carcinoma SGC-996 cells was detected by colony formation assay. The apoptosis and cell cycle distribution of gallbladder carcinoma SGC-996 cells after treatment with 40, 60 and 80 μg/mL patchoulenone for 48 h were detected by FCM. The expression levels of cleaved caspase-9, cleaved caspase-3, cleaved ploy （ADP-ribose） ployrnerase-1 （PARP-1）, Bax, Bcl-2, cyclin D1, cyclin A and cyclin B1 proteins in gallbladder carcinoma SGC-996 cells after treatment with 40, 60 and 80 μg/mL patchoulenone for 48 h were detected by Western blotting. Results： Patchoulenone inhibited the proliferation of gallbladder carcinoma SGC-996 cells after treatment with 10, 40, 80, 100 and 120 μg/mL patchoulenone （all P 〈 0.01）, and in a dose and time-dependent manner （all P 〈 0.05）. Patchoulenone inhibited the colony formation of human gallbladder carcinoma SGC-996 cells after treatment with 10, 20 and 30μg/mL patchoulenone （alJ P 〈 0.05）. Patchoulenone significantly induced the apoptosis of SGC-996 cells after treatment with 40, 60 and 80 μg/mL patchoulenone and led to S-phase cell-cycle arrest （all P 〈 0.05）. The expression levels of cleaved caspase-9, cleaved caspase-3, cleaved PARP-1 protein and the ratio of Bax to Bcl-2 in SGC-996 cells after treatment with 40, 60 and 80 μg/mL were up-regulated （all P 〈 0.05）, whereas the expression levels of cyclin D1, cyclin A and cyclin B1 were down-regulated （all P 〈 0.01）. Conclusion： Patchoulenone can inhibite the proliferation of human gallbladder carcinoma SGC-996 cells and arrest the cell cycle at

关 键 词：胆囊肿瘤 细胞增殖 细胞凋亡 广藿香酮 

分 类 号：R735.8[医药卫生—肿瘤]