野生型RET与RET／PTC融合基因在甲状腺乳头状癌的表达

Expression of wild-type RET and RET/PTC fusion gene in papillary thyroid carcinoma

目的探讨野生型RET（wildtype RET，WT—RET）和RET／胛C融合基因的表达与安徽地区人群胛C发生、发展的关系。方法收集新鲜的35例胛C和相应的35例癌旁组织及10例甲状腺良性病变旁正常组织，巢氏PCR检测RET／阿C1、2和3的表达，DNA测序检测融合基因序列。实时荧光定量PCR检测WT．RET和RET酪氨酸激酶区（RETtyrosinekinase，RET．TK）mRNA表达水平。原位杂交法检测RET的表达位置。结果35例PTC中，RET／PTC阳性组织17例，其中RET／PTCl阳性15例，RE唧C2阳性0例，RET，PTC3阳性2例，并运用DNA测序证实。WT．RET与RET．TK2者在癌旁组织都几乎不表达，2者在RET／PTC阴性组的表达都是各自癌旁组织的2倍（P〈0．01，P〈0．01），2者在RE删P1rC阳性组的表达分别是各自癌旁组织的3倍（P〈0．01）和64倍（P〈0．01）。WT—RET在RE删P1rC阳性与阴性2组之间表达差异无统计学意义（乃O．05），RET-TK在RET／PTC阳性组表达明显高于阴性组，差异有统计学意义（P〈O．01）。原位杂交结果显示RET表达于PTC滤泡细胞膜和细胞浆。RE唧C与PTC临床参数如性别、年龄、肿瘤直径、淋巴结转移及临床分期等均差异无统计学意义（P〉0．05）。结论①安徽地区人群散发性PTC中RET／PTC表达率不高，其对PTC的诊断和判断预后的价值不大。②散发性PTC中滤泡细胞表达WT—RET，可能促进了RET／PTC的发生，并共同参与了甲状腺肿瘤的发生。

Objective To explore the relationship between wild type RET （WT-RET）and RET/PTC fusion gene expression and the development of papillary thyroid carcinoma（PTC）. Methods 35 cases of fresh PTC tissues, the corresponding paracancerous thyroid tissues and 10 cases of fresh adjacent benign thyroid lesions normal tissues were collected. Expression of RET/PTC 1, 2 and 3 fusion gene were detected by Nested PCR and DNA sequence test. Expression of WT-RET and RET tyrosine kinase （RET-TK）mRNA were detected by quantitative Real Time PCR. RET mRNA expression in the source was detected by in situ hybridization. Results A- mong the 35 cases of PTC,17 cases had positive RET/PTC fusion gene（15 cases of positive RET/PTC 1, 0 case of positive RET/PTC 2 was, 2 cases of positive RET/PTC 3）, and all positive cases were confirmed by DNA sequence test. Almost no expression of WT-RET and RET-TK mRNA was found in paracancerous thyroid tissues, while their expressions in RET/PTC negative group were 2 times higher than those in the respective paracancerous thyroid tissues （P〈0.01, P〈0.01 ）and their expression in RET/PTC positive group was 3 times and 64 times higher than the respective paracancerous thyroid tissues （P〈0.01, P〈0.01 ）.Expression of WT-RET mRNA had no statistical difference between RET/FTC positive group and negative group （P〉0.05）. Expression of RET-TK mRNA in RET/PTC positive group was significantly higher than that in the negative group, and the difference has statistical significance （P〈0.01）. In situ hybridization showed RET mRNA expressed in the cell membrane and cytoplasm. Expression of RET/PTC fusion gene was not statistically correlated with clinical parameters such as age, gender, tumor size, lymph node metastasis or clinical stage（P〉0.05）. Conclusions ①Expression ratio of RET/ PTC positive is not high in sporadic PTC in Anhui province,and its diagnostic and prognosis significance is limited. ②Expression of WT RET in follicular cells in sporadic FFC may promote the occurance of RET/FFC fusion gene, and the two may commonly involved in the occurrence of thyroid tumor.