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中国癌症防治杂志

中国癌症防治杂志 ›› 2014, Vol. 6 ›› Issue (1): 7-11.doi: 10.3969/j.issn.1674-5671.2014.01.02

• 基础研究 • 上一篇    下一篇

Axin-1在C57BL/6J小鼠肝癌模型发生过程中的表达及意义

  

  1. 广西肿瘤防治研究所实验研究部；广西医科大学研究生学院
  • 出版日期:2014-03-25 发布日期:2014-04-10
  • 通讯作者: 匡志鹏 E-mail:kgzhou@163.com
  • 基金资助:

    国家自然科学基金资助项目（30760278）

Expression and significance of Axin-1 during chemical induction of liver cancer in C57BL/6J mice

  • Online:2014-03-25 Published:2014-04-10

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被引次数

6

摘要： 目的 检测Wnt/β-catenin信号通路中Axin-1在化学诱导C57BL/6J小鼠肝癌模型发生过程中的表达水平，揭示Wnt/β-catenin通路与肝癌发生的关系。方法 95只C57BL/6J小鼠随机分为实验组50只和对照组45只。实验组小鼠以联合二乙基亚硝胺（DEN）/四氯化碳（CCl4）/乙醇诱导建立小鼠肝癌模型，对照组未予任何特殊处理。每2周定期处死两组小鼠各5只，并取肝组织进行病理学观察，以实时荧光定量PCR（RT-PCR）技术、Western blot法、免疫组化法检测并动态观察Axin-1 mRNA和蛋白水平的表达情况。结果 小鼠经化学诱导20周后成功诱发小鼠肝癌并建立小鼠肝癌模型。RT-PCR和Western blot法检测显示，Axin-1 mRNA和蛋白水平的表达在诱导的第4周至第14周，实验组和同期对照组比较差异无统计学意义（P ＞0.05）；在诱导的第16周至第20 周实验组和同期对照组比较差异有统计学意义（P＜0.05），且随着诱癌时间的延长，实验组Axin-1的表达逐渐降低，在诱导的第16周、第18周和第20周Axin-1 mRNA的相对表达量分别为0.421±0.083、0.278±0.042、0.120±0.028（P＜0.05）。免疫组化法检测显示Axin-1在对照组各诱导期均为阳性表达，实验组从第16 周开始Axin-1的表达减弱，至第 20周时Axin-1几乎不表达。结论 Wnt/β-catenin信号通路可能与肝癌的发生、发展有关，其中Axin-1可能起着负性调节的作用。

关键词: 肝肿瘤, Wnt/β-catenin 信号通路, Axin-1, 化学诱癌

Abstract: Objective To analyze the expression of Axin-1 during chemical induction of liver cancer in C57BL/6J mice. Methods C57BL/6J mice were divided randomly into an experimental group（n=50）and a control group（n=45）.Mice in the experimental group were treated with diethylnitrosamine（DEN），carbon tetrachloride（CCl4） and ethanol for 20 weeks to induce hepatocellular carcinoma （HCC）.Every 2 weeks，5 mice in each group were sacrificed and liver samples were taken.Dynamic expression of Axin-1 was analyzed using real-time PCR，Western blotting and immunohistochemistry，while pathological changes were analyzed using hematoxylin staining. Results The 20-week DEN/CCl4/ethanol treatment successfully induced liver cancer in C57BL/6J mice.Based on real-time PCR and Western blotting，expression of Axin-1 mRNA and protein was significantly lower in the experimental group than in the control group between weeks 16 and 20.This expression decreased gradually over time in the experimental group：levels of mRNA were 0.421±0.083 at week 16，0.278±0.042 at week 18 and 0.120±0.028 at week 20（P＜0.05）.Immunohistochemistry showed detectable levels of Axin-1 staining in the control group at all time points tested；in the experimental group，however，the level of staining began to decrease at week 16 and was nearly undetectable at week 20. Conclusion The Wnt/β-catenin signaling pathway may be associated with HCC development，with Axin-1 acting to inhibit HCC progression.

Key words: Liver neoplasm, Wnt/β-catenin, Signaling pathway, Axin-1, Chemical carcinogenesis