CAJ | 학술논문

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。
목적：검측Wnt/β-catenin신호통로중Axin-1재화학유도C57BL/6J소서간암모형발생과정중적표체수평，게시Wnt/β-catenin통로여간암발생적관계。방법95지C57BL/6J소서수궤분위실험조50지화대조조45지。실험조소서이연합이을기아초알（DEN）/사록화탄（CCl4）/을순유도건립소서간암모형，대조조미여임하특수처리。매2주정기처사량조소서각5지，병취간조직진행병이학관찰，이실시형광정량PCR（RT-PCR）기술、Western blot법、면역조화법검측병동태관찰Axin-1 mRNA화단백수평적표체정황。결과소서경화학유도20주후성공유발소서간암병건립소서간암모형。RT-PCR화Western blot법검측현시，Axin-1 mRNA화단백수평적표체재유도적제4주지제14주，실험조화동기대조조비교차이무통계학의의（P>0.05）；재유도적제16주지제20주실험조화동기대조조비교차이유통계학의의（P<0.05），차수착유암시간적연장，실험조Axin-1적표체축점강저，재유도적제16주、제18주화제20주Axin-1 mRNA적상대표체량분별위0.421±0.083、0.278±0.042、0.120±0.028（P<0.05）。면역조화법검측현시Axin-1재대조조각유도기균위양성표체，실험조종제16주개시Axin-1적표체감약，지제20주시Axin-1궤호불표체。결론 Wnt/β-catenin신호통로가능여간암적발생、발전유관，기중Axin-1가능기착부성조절적작용。
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.