甲状腺过氧化物酶mRNA cRNA探针的构建及意义  

Synthesis of thyroperoxidase mRNA cRNA probe and its application

作　　者：陈云[1] 朱明华[2] 周晓军[3] 倪灿荣[2] 

机构地区：[1]解放军第411医院病理科,上海200081 [2]第二军医大学长海医院病理科,上海200433 [3]南京军区南京总医院病理科,南京210002

出　　处：《临床与实验病理学杂志》2008年第5期584-588,共5页Chinese Journal of Clinical and Experimental Pathology

摘　　要：目的探讨甲状腺过氧化物酶(thyroperoxidase,TPO)mRNA cRNA探针的构建及原位表达的检测方法和意义。方法取甲状腺结节新鲜组织,在RT-PCR扩增TPO cDNA的基础上,构建pSPT19-TPO质粒,经HindⅢ和BamHⅠ单酶切后成线性化模板,在SP6和T7 RNA聚合酶作用下,体外转录合成地高辛标记的TPO cRNA反义和正义探针,进行TPO mRNA的原位杂交实验。结果TPO mRNA阳性杂交信号分布于甲状腺滤泡细胞的胞质,本组腺瘤2例、结节性甲状腺肿4例、瘤周甲状腺组织1例原位杂交阳性,乳头状癌2例、桥本病1例原位杂交阴性。核酸原位杂交和RT-PCR检测甲状腺组织TPO mRNA的表达,结果一致。结论成功构建甲状腺过氧化物酶mRNA的cRNA探针;检测TPO mRNA是一种较准确反映甲状腺组织TPO状态的方法,TPO mRNA的原位检测可结合组织形态学了解甲状腺滤泡细胞的功能表达状况。Purpose Synthesis of thyroperoxidase mRNA cRNA probe and its application to In Situ Hybridization.Methods Fresh thyroid tissues originated from thyreoidectomy were obtained.On the basis of TPO cDNA amplified with RT-PCR technique,the plasmid of pSPT19-TPO was constituted.And then pSPT19-TPO was separately digested by Hind Ⅲ and BamHⅠenzyme to form the linearization of expression plasmia.TPO cRNA sense and antisense probes were labeled with digxigenin in vivo by using SP6 and T7 RNA polymerase so that TPO mRNA tested by in situ hybridization histochemistry technique（ISHH） was carried out in thyroid tissues.Results The positive signal of TPO mRNA tested by ISHH was found in the cytoplasm of thyroid follicular cell.The positive result was detected in two cases of adenoma,four cases of nodular goiter and peri-neoplasm thyroid tissues.The two cases of papillary carcinoma and one Hashimoto′ disease were showed negative results.The TPO mRNA expression in both of RT-PCR and ISHH was in accordance with each other.Conclusion Thyroperoxidase mRNA cRNA probe was made successfully.To test TPO mRNA is one of the accurate methods to represent the expression of thyroperoxidase in thyroid tissue.In situ detecting thyroperoxidase mRNA may combine with histomorphology so as to understand the functional expressive states of thyroid follicular cells.

关 键 词：甲状腺过氧化物酶 甲状腺 CRNA探针 原位杂交 

分 类 号：R345.46[医药卫生—基础医学]