Zebularine对胆管癌细胞增殖、凋亡的影响及其机制  

Effect of Zebularineon cell proliferation and apoptosis in QBC939 cholangiocarcinoma cells and its mechanisms

作　　者：熊辉[1] 丁佑铭[1] 廖晓峰[2] 

机构地区：[1]武汉大学人民医院肝胆腹腔镜外科,湖北武汉430060 [2]襄阳市中心医院普外科

出　　处：《胃肠病学和肝病学杂志》2015年第7期864-867,共4页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的观察新型DNA甲基转移酶抑制剂Zebularine对胆管癌细胞系QBC939细胞增殖、凋亡的影响,并分析其可能的机制。方法体外培养的QBC939细胞,经不同浓度Zebularine(0、50、100、200μmol/L)分别处理24、48、72 h,采用MTT法检测其对细胞增殖的影响;Zebularine处理QBC939细胞48 h后,流式细胞仪检测其对细胞凋亡的影响;分光光度法检测药物处理后细胞Casepase-9相对活性;Western blotting法检测药物对细胞Bax、Bcl-2、PTEN、Akt、p-Akt蛋白表达的影响。结果 Zebularine能有效抑制QBC939细胞增殖,且呈剂量和时间依赖性(P<0.05);Zebularine处理QBC939细胞48 h后,细胞凋亡率从16.7%增加到37.5%;促凋亡蛋白Bax表达量上调,而抗凋亡蛋白Bcl-2表达量下调(P<0.05);Casepase-9相对活性显著增加(P<0.05);此外,Zebularine可上调PTEN表达,并抑制Akt磷酸化(P<0.05)。结论 Zebularine可通过抑制QBC939细胞增殖并促进其凋亡,其作用机制可能参与上调PTEN表达并抑制Akt磷酸化激活,继而上调Bax,下调Bcl-2,并增加Caspase-9活性相关。Objective To investigate the effects of Zebularine on the cell proliferation and apoptosis in cholangiocarcinoma cell line QBC939 cells and explore the possible mechanisms. Methods QBC939 cells were cultured in vitro.After treatment by Zebularine in different concentrations respectively at different time,the cell survival was determined by the MTT method. Apoptosis was detected by flow cytometry. The relative activity of Caspase-9 was monitored by spectrophotometer. Western blotting was used for Bcl-2,Bax,PTEN,Aktand p-Aktprotein analysis. Results The cell proliferation of QBC939 cells was inhibited by Zebularine in a dose-dependent and time-dependent manner（ P〈0. 05）.Being treated with Zebularine after 48 hours,the apoptosis rate of QBC939 cells increased from 16. 7% to 37. 5%. Zebularine promoted the expression of pro-apoptotic factor Bax,and suppressed the expression of anti-apoptotic factor Bcl-2 in a dose-dependent manner（ P〈0. 05）. The relative activity of Caspase-9 of Zebularine group was increased（ P〈0. 05）. Furthermore,the data of Western blotting showed that Zebularine decreased the expression of p-Akt in a dosedependent manner（ P〈0. 05）. Conclusion Zebularine inhibits the cell proliferation and induces cell apoptosis in QBC939 cells,which is associated with the modulation of expressions of PTEN and Akt,thus up-regulating of Bax,down-regulating of Bcl-2 and activating of Caspace-9.

关 键 词：ZEBULARINE QBC939细胞 凋亡 PTEN AKT 

分 类 号：R735.8[医药卫生—肿瘤]