细针穿刺标本及外周循环中miR-191和miR-494对甲状腺癌的早期诊断价值

Early diagnostic value of miR-191 and miR-494 from fine needle aspiration specimens and peripheral circulation for thyroid carcinoma

目的 探讨良恶性甲状腺结节中死亡相关蛋白激酶（DAPK）相关miR-191和磷酸酶-张力蛋白基因（PTEN）相关miR-494在细针穿刺细胞学检查（FNAB）标本、外周循环中的表达水平及差异,为良恶性甲状腺结节疾病的鉴别提供分子诊断依据.方法 收集113例甲状腺结节（恶性及可疑恶性甲状腺结节48例、结节性甲状腺肿38例、甲状腺腺瘤27例）患者的FNAB标本及外周静脉血,应用实时荧光定量逆转录聚合酶链反应（qRT-PCR）检测其中98例甲状腺结节患者FNAB标本及外周循环中miR-191和miR-494的表达水平.通过受试者工作特征曲线（ROC曲线）分析miR-191和miR-494,诊断恶性甲状腺结节的灵敏度和特异度.结果 （1）FNAB诊断结果与手术病理结果诊断的符合率为91.35％,FNAB诊断甲状腺癌的灵敏度为91.7％（44/48）,特异度为90.9％（30/33）.（2）FNAB标本及外周循环中,甲状腺癌组miR-191的表达水平明显低于良性组,而甲状腺癌组miR-494表达水平明显高于良性组（P〈0.05）.（3）FNAB标本及外周循环中,miR-191和miR-494对恶性甲状腺结节的诊断灵敏度与特异度均尚可（ROC曲线下面积〉0.7）.miR-191在FNAB标本和外周循环的灵敏度、特异度分别为76.9％、73.5％和61.5％、64.1％;miR-494分别为63.6％、76.5％和72.7％、84.6％.（4）在FNAB标本及外周循环中,甲状腺癌中miR-191和miR-494的相对表达量与年龄、性别、结节大小、有无钙化、有无颈部淋巴结肿大、甲状腺功能及甲状腺抗体有无异常这些临床基本特征之间的差异没有统计学意义（P〉0.05）.结论 miR-191和miR-494可作为辅助FNAB早期诊断甲状腺癌的分子标记物.

Objective To investigate the different expression levels of death-associated protein kinase （ DAPK） related miR-191 and phosphatase and tensin homolog deleted on chromosome ten （ PTEN） related miR-494 from fine-needle aspiration biopsy （ FNAB） samples and blood of both benign and malignant thyroid nodules, and to find new clinical molecular diagnostic markers. Methods FNAB specimens and peripheral venous blood were collected from 113 patients with thyroid nodules （48 cases of malignant and suspected malignant thyroid nodules, 38 cases of nodular goiter, and 27 cases of thyroid adenomas）. The expression levels of miR-191 and miR-494 were detected by realtime fluorescence quantitative reverse transcription PCR （ qRT-PCR ） . qRT-PCR were applied to detect miR-191 and miR-494 expression level in 98 patients with thyroid nodules and peripheral circulation. Receiver operating characteristic curves （ ROC curves） were used to evaluate the sensitivity and specificity of miR-191 and miR-494 to diagnose malignant thyroid nodules. Results （1） The sensitivity of FNAB in diagnosing thyroid cancer was 91. 7% （44/48） and the specificity was 90. 9% （30/33）,the diagnostic accuracy was 91. 35%. （2） In FNAB samples, as well as in peripheral circulation, the relative expression of miR-191 in thyroid cancer group is significantly lower than that of the benign group, while the relative expression of miR-494 in thyroid cancer group is significantly higher than that of the benign group （P〈0. 05）. （3） The sensitivity and specificity of miR-191 and miR-494 were acceptable （area under the ROC curve〉0. 7）. Sensitivity and specificity of miR-191 in FNAB and peripheral circulation were 76. 9%, 73. 5% and 61. 5%, 64. 1%; miR-494 were 63. 6%,76. 5% and 72. 7%, 84. 6%respectively. （4） In thyroid cancer FNAB samples and peripheral circulation, the differences between the relative expression level of miR-191 and miR-494, and the clinical characteristics of age, gender, nodule size, and calcification, with or without cervical lymph node enlargement, thyroid function and thyroid antibodies with or without abnormalities were not statistically significant（P〉0. 05）. Conclusion MiR-191 and miR-494 can be used as molecular diagnostic markers for early diagnosis of thyroid carcinoma with adjunctive FNAB.