Axin-1 在 C57BL/6J 小鼠肝癌模型发生过程中的表达及意义

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。

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

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 （CCL） 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/CCh/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 significant/y 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）.Immunohistoehemistry 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 undeteetable at week 20. Conclusion The Wnt/β-catenin signaling pathway may be associated with HCC development, with Axin-1 acting to inhibit HCC progression.