蟾毒灵抑制胆总管癌细胞QBC-939增殖和诱导凋亡的实验研究  

Anticancer effect of Bufalin on suppression of cell proliferation and induction of cell apoptosis in cholangiocellular carcinoma QBC-939 cells

作　　者：郑亦胡[1,2] 胡伟建[3] 赵炜嘉 余华军[1] 张启瑜[1] ZHENG Yi-hu;HU Wei-jian;ZHAO Wei-jia;YU Hua-jun;ZHANG Qi-yu(Department of Hepatobiliary Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China;Derpartment of General Surgery, Yongjia Chinese Medicine Hospital, Wenzhou, Zhejiang 325401, China;Department of General Surgery, Jinhua Municipal Central Hospital, Jinhua, Zhejiang 321000, China)

机构地区：[1]温州医科大学附属第一医院肝胆外科,浙江温州325000 [2]浙江省永嘉县中医医院普外科,浙江温州325401 [3]金华市中心医院普外科,浙江金华321000

出　　处：《肝胆胰外科杂志》2017年第3期232-236,共5页Journal of Hepatopancreatobiliary Surgery

基　　金：温州市中医药科学研究资助项目(2016ZB08)

摘　　要：目的观察蟾毒灵对人胆管癌细胞QBC-939增殖和凋亡的影响,以及凋亡相关蛋白表达量的变化。方法体外培养人胆管癌QBC-939细胞,运用MTT检测蟾毒灵对胆管癌细胞增殖的影响,采用流式细胞仪检测蟾毒灵对胆管癌细胞凋亡率和线粒体膜电位的影响;并采用Western blotting检测凋亡相关蛋白Caspase3、激活型的Caspase3(Cleaved-Caspase3)和Cleaved-PARP的表达量变化。结果蟾毒灵对人胆管癌细胞QBC-939增殖具有明显抑制作用,在一定浓度下表现出时间、剂量依赖性。流式细胞仪检测显示:细胞的凋亡率明显增加;线粒体膜电位随蟾毒灵浓度增加,膜电位发生下调。Western blotting结果显示:蟾毒灵处理QBC-939细胞48h后,Caspase3蛋白被剪接活化,激活型的Caspase3(Cleaved-Caspase3)表达量升高;同时,Caspase3的作用底物多聚聚合酶PARP的降解产物(Cleaved-PARP)表达明显升高,且呈浓度依赖性。结论蟾毒灵能够明显抑制人胆管癌QBC-939细胞的增殖,并呈时间和浓度依赖性;线粒体凋亡途径在蟾毒灵的抗癌过程中发挥了作用。Objective To investigate the effects of Bufalin on cell proliferation,apoptosis and the changes of apoptosic related proteins in cholangiocellular carcinoma cell line QBC-939.Methods The human cholangiocarcinoma cell line QBC-939was cultured in vitro.Cell viability was measured by MTT assay.The apoptotic rate and the mitochondrial membrane potential△Ψm were analyzed by flow cytometry analysis.The apoptotic protein level of Caspase3,Cleaved-Caspase3and Cleaved-PARP were detected by Western blotting.Results Bufalin had a significant inhibitory effect on the growth of cholangiocarcinoma cell line QBC-939in a time-and dosedependent manner.The result of FCM indicated that the apoptotic rate of cultured cell line QBC-939increased under Bufalin treatment.Moreover,Western blotting results showed enhanced levels of both Cleaved-Caspase3and Cleaved-PARP expression in a dose dependent manner,which suggested that Bufalin could stimulate Caspase3activation.In contrary,the mitochondrial membrane potential△Ψm reduced with increasing concentration of Bufalin.Conclusion Bufalin has a prominent anti-proliferation effect on cholangiocarcinoma cell line QBC-939in a time-and dose-dependent manner.Bufalin may exert its role as an anticancer regime through surrogating the mitochondrial apoptosis pathway in QBC-939cells.

关 键 词：蟾毒灵 人胆管癌细胞QBC-939 凋亡 增殖 

分 类 号：R285[医药卫生—中药学]