Regulation on expression of Gankyrin by hepatitis B virus X protein through Wnt/β-catenin signaling

Liu Rui; Wei Xufu; Shen Ai; Wu Zhongjun

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Regulation on expression of Gankyrin by hepatitis B virus X protein through Wnt/β-catenin signaling

更新时间：2023-12-18

- Regulation on expression of Gankyrin by hepatitis B virus X protein through Wnt/β-catenin signaling
- Journal of Chongqing Medical University Vol. 38, Issue 7, Pages: 923-928(2014)
- 作者机构：
  
  重庆医科大学附属第一医院肝胆外科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2014，
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Liu Rui, Wei Xufu, Shen Ai, et al. Regulation on expression of Gankyrin by hepatitis B virus X protein through Wnt/β-catenin signaling. [J]. Journal of Chongqing Medical University 38(7):923-928(2014)
DOI：

Liu Rui, Wei Xufu, Shen Ai, et al. Regulation on expression of Gankyrin by hepatitis B virus X protein through Wnt/β-catenin signaling. [J]. Journal of Chongqing Medical University 38(7):923-928(2014) DOI：

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。

Abstract

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.

关键词

肝癌乙肝病毒X蛋白Gankyrinβ-catenin

Keywords

hepatocellular carcinomahepatitis B virus X pro－teinGankyrinβ-catenin

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