Molecular analysis of thyrotropin receptor in thyroid noeplasm

从34例甲状腺组织（4例正常、6例腺瘤、6例滤泡癌和18例乳头状癌）中提取总RNA，反转录成单链cDNA，然后将其作为聚合酶链反应的模板，进行单链构象多态性（SSCP）分析。两个片段FRAG（468-692）（核苷酸2044 - 2295，对应于TSH-R cDNA的COOH-末端，胞质末端）在不同甲状腺组织样品中的电泳迁移率存在差异。直接测序显示FRAG（468-692）中Phe^<197>（TTC）* Ile（ATC）和Asp^<219>（GAT）* Glu（GAG）取代形成两种乳头状癌。在FRAG（2044-2295）中鉴定了三种类型的取代：<715>来自一种乳头状癌的Asn^（AAC）* Asp（GAC），<723>来自一种乳头状癌的Lys^（AAG）* Met（ATG），以及<727>来自一种正常组织样品、一种滤泡癌和四种乳头状癌的Asp^（GAC）* Glu（GAG）。提示部分甲状腺肿瘤组织中存在TSH-R结构的改变。

Total RNA was prepared from 34 thyroid tisssue samples (four normal, six adenoma, six follicular cancer, and 18 papillary cancer) and reverse-transcribed into single-stranded cDNA,which was then used as a templated for the polymerase chain reaction and subjected to single-strand conformation polymorphism (SSCP) analysis. Two fragments, FRAG (468-692) (nucleotides 2044 to 2295, corresponding to the COOH-terminal, cytoplasmic dmain of the TSH-R cDNA) showed differences in electrophoretic mobility among the various thyroid tissue samples. Direct sequencing revealed Phe^<197> (TTC) * Ile (ATC), and Asp^<219> (GAT) * Glu (GAG) substitutions in FRAG (468-692) form two papillary cancers. Three types of substitution were identified in FRAG (2044-2295) : Asn^<715> (AAC) * Asp (GAC) from one papillary cancer, Lys^<723> (AAG) * Met (ATG) from one papillary cancer, and Asp^<727> (GAC) * Glu (GAG) from one normal tissue sample, one follicular cancer, and four papillary cancers. These results suggest that there exist structure changes in TSH-R in some cases of thyroid neoplastic tissue.