引用本文:刘 锐,魏续福,沈 艾,吴忠均.乙肝病毒X蛋白通过Wnt/β-catenin信号通路调控Gankyrin表达[J].重庆医科大学学报,2014,38(7):923~928
乙肝病毒X蛋白通过Wnt/β-catenin信号通路调控Gankyrin表达
Regulation on expression of Gankyrin by hepatitis B virus X protein through Wnt/β-catenin signaling
DOI:
中文关键词:  肝癌  乙肝病毒X蛋白  Gankyrin  β-catenin
英文关键词:hepatocellular carcinoma  hepatitis B virus X pro-tein  Gankyrin  β-catenin
基金项目:
作者单位
刘 锐,魏续福,沈 艾,吴忠均 重庆医科大学附属第一医院肝胆外科重庆 400016 
摘要点击次数: 726
全文下载次数: 435
中文摘要:
      目的:研究乙肝病毒X蛋白(hepatitis B virus X protein,HBX)对Gankyrin表达水平的影响以及Wnt/β-catenin信号通路在HBX基因调控Gankyrin表达中的作用。方法:HBX腺病毒表达载体(Ad-HBX-GFP)感染转染2种人肝癌细胞系HepG2、SMMC-7721细胞,Si-HBX分子(Si-HBX oligo)用 LipofectamineTM 2000转染入HepG2.2.15细胞,用RT-PCR和Western blot法检测HBX与Gankyrin的表达情况;β-catenin腺病毒载体转染HepG2、SMMC-7721细胞,用Western blot法检测β-catenin与Gankyrin的表达情况;在沉默β-catenin表达的HepG2、SMMC-7721细胞中过表达HBX基因,Western blot法检测β-catenin与Gankyrin的表达情况。结果:①HepG2.2.15细胞中Gankyrin mRNA与蛋白表达水平均高于HepG2细胞(P<0.05)。②HepG2与SMMC-7721细胞系中Ad-HBX-GFP组Gankyrin mRNA与蛋白表达水平均高于绿色荧光蛋白腺病毒表达载体(Ad-GFP)组(P<0.01)。③HepG2.2.15细胞中Si-HBX组Gankyrin mRNA与蛋白表达水平均低于Si-阴性对照(Si-NC)组(P<0.01)。④在HepG2与SMMC-7721细胞系中过表达HBX后,Ad-HBX-GFP组β-catenin的蛋白表达水平均高于Ad-GFP组(P<0.01)。⑤在HepG2与SMMC-7721细胞系中过表达β-catenin后,β-catenin腺病毒表达载体(Ad-β-catenin-GFP)组Gankyrin的蛋白表达水平均高于Ad-GFP组(P<0.01)。⑥在HepG2与SMMC-7721细胞系中沉默β-catenin后,过表达HBX基因,Si-β-catenin腺病毒表达载体(Ad-si-β-catenin-RFP)组与红色荧光蛋白腺病毒表达载体(Ad-RFP)组相比,Gankyrin的蛋白表达水平未见升高(P<0.01)。结论:HBX-Wnt/β-catenin-Gankyrin信号转导通路存在与肝癌细胞中,抑制β-catenin的功能可阻遏HBX对Gankyrin的上调作用,本结果将可能为肝癌的治疗提供新的思路。
英文摘要:
      Objective:To investigate the effects of HBX on Gankyrin expression and role of Wnt/β-catenin signaling pathway in the regulation of HBX on Gankyrin. Methods:HBX adenovirus vector Ad-HBX-GFP was transfected into human hepatocellular carcinoma cell line HepG2,SMMC-7721 cells and Si-HBX molecules was transfected into HepG2 2.15 cells with LipofectamineTM 2000. Expres-sions of HBX and Gankyrin were detected by RT-PCR and Western blot. β-catenin adenovirus vector was transfected into HepG2 and SMMC-7721 cells,and expressions of β-catenin and Gankyrin were detected by Western blot. HBX was overexpressed in β-catenin silenced HepG2 and SMMC-7721 cells,and expressions of β-catenin and Gankyrin were detected by Western blot. Results:①Expres-sions of Gankyrin were higher in HepG2.2.15 cells than in HepG2 cells(P<0.05). ②mRNA and protein expressions of Gankyrin were increased significantly in Ad-HBX-GFP group than in Ad-GFP group in HepG2 and SMMC-7721 cell line(P<0.01). ③mRNA and protein expressions of Gankyrin were decreased significantly in Si-HBX group than in Si-NC group in HepG2.2.15 cells(P<0.05). ④Protein expressions of β-catenin were increased significantly in Ad-HBX-GFP group than in Ad-GFP group after overexpressing HBX protein in HepG2 and SMMC-7721 cell line(P<0.01). ⑤Protein expressions of Gankyrin were increased significantly in Ad-HBX-GFP group than in Ad-GFP group after overexpressing β-cateninin HepG2 and SMMC-7721 cell line(P<0.01). ⑥Protein ex-pressions of Gankyrin were not increased in Ad-si-β-catenin-RFP group than in Ad-RFP group after silencing β-catenin in HepG2 and SMMC-7721 cell line and overexpressing HBX gene(P<0.01). Conclusion:HBX-Wnt/β-catenin-Gankyrin signal transduc-tion pathways are existed in hepatocarcinogenesis. Inhibiting the function of β-catenin can interrupt the up-regulation of Gankyrin by HBX,which may provide a new treatment method for hepa-tocellular carcinoma.
查看全文  查看/发表评论  下载PDF阅读器
关闭
微信关注二维码