摘要
目的探讨C57BL/6J小鼠诱癌过程中Hedgehog信号通路成员Shh、Ptch和Smo在各阶段的动态表达。方法将95只正常C57BL/6J雄性小鼠随机分为诱癌组50只(化学诱癌处理)和对照组45只(正常喂养),定期获取小鼠肝组织标本行免疫组织化学法、实时荧光定量RT-PCR法和蛋白质印迹法检测。结果免疫组织化学提示,诱癌组Shh、Ptch和Smo蛋白分别在第6周、第8周和第10周出现弱阳性,均在第16周出现中度阳性,第20周出现强阳性表达,而正常对照组中未检测出三者的表达。RT-PCR法提示,诱癌组小鼠肝组织中Shh mRNA、Ptch mRNA和Smo mRNA的表达量持续升高,第20周时三者的表达水平均达到最高(Shh mRNA 15.986±1.784,Ptch mRNA 11.272±0.295,Smo mRNA 10.185±0.716),明显高于同期正常对照组(P<0.05)。蛋白质印迹法提示,诱癌组Shh、Ptch和Smo蛋白分别从第6周、第8周和第10周起出现弱表达,到第20周时显著增强,而正常对照组中未检测出3种蛋白的表达。结论 Hedgehog信号通路分子Shh、Ptch和Smo伴随着小鼠诱发肝癌时间的延长其表达量逐步升高,认为Hedgehog信号通路与小鼠肝癌的发生、发展密切相关。
Objective To explore Hedgehog signaling pathway members Shh,Ptch and Smo dynamic expression in each stage of in- duced C57BL/6J mice liver cancer process. Methods 95 normal C57BL/6J male mice were randomly divided into chemical - induced group（50 mice） and normal control group （45 mice）. Liver tissue was regularly obtained to detect by immunohistochemistry, RT -PCR and western blot. Results Immunohistoehemistry showed Shh,Ptch and Smo expression of the chemical - induced group were weakly pos- itive on the 6th,thc 8th and the 10th week. Then all of three appeared moderate expression at 16th week and strong expression at 20th week. While the normal control group was not detected for them. RFQ - PCR showed Shh mRNA,Pteh mRNA and Smo mRNA expression of the chemical -induced group continued to rise,and at the 20th week all of three expression had reached the highest level（ Shh mRNA was 15. 986 ± 1. 784, Ptch mRNA was 11. 272 ± 0. 295, Smo mRN A was 10. 185 ±0.716）, and were significantly higher than the normal control group （P 〈 0.05 ）. Western - blot showed the Shh, Ptch and Smo protein expression of the chemical - induced group appeared weak expression from the 6th,the 8th and the 10th week. At the 20th week,all proteins expression enhanced significantly. While the normal con- trol group were not detected for them. Conclusion Accompanied by the time prolonged,Hedgehog signaling pathway gene Shh,Pteh and Smo expression level had been elevated,and is closely related with the occurrence and development of hepatocellular carcinoma in mice.
出处
《医学研究杂志》
2014年第5期65-69,共5页
Journal of Medical Research
基金
广西研究生科研创新基金资助项目(2011105981002M180)
