评估血浆外泌体 miRNA 作为甲状腺乳头状癌淋巴结转移的潜在诊断生物标志物。

Evaluation of plasma exosomal miRNAs as potential diagnostic biomarkers of lymph node metastasis in papillary thyroid carcinoma.

机构信息

Thyroid Surgery Center, West China Hospital of Sichuan University, Chengdu, 610041, China.

Laboratory of Thyroid and Parathyroid Disease, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.

出版信息

Endocrine. 2022 Mar;75(3):846-855. doi: 10.1007/s12020-021-02949-x. Epub 2021 Dec 1.

DOI:10.1007/s12020-021-02949-x

PMID:34854020

Abstract

PURPOSE

The early diagnosis of lymph node metastasis (LNM) in papillary thyroid carcinoma (PTC) is clinically important, as it can aid in treatment decision-making and improve prognosis. In the present study, we aimed to identify whether plasma exosomal miRNAs could be potential diagnostic markers of LNM in PTC.

METHODS

Profiles of plasma exosomal miRNAs were screened using miRNA microarrays. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed in the validation and diagnostic sets to select candidate exosomal miRNAs. Finally, receiver operating characteristic (ROC) curves were generated to evaluate the efficiency of target exosomal miRNAs in distinguishing PTC-N1 patients from PTC-N0 patients.

RESULTS

In total, 197 miRNAs were found to be differentially expressed in the testing set. Based on the qRT-PCR results, the expression of miR-6774-3p (p < 0.001) and miR-6879-5p (p < 0.001) in the PTC-N1 patients was significantly higher than that in the controls. The AUC values of plasma exosomal miR-6774-3p (0.802; 95% CI, 0.724-0.879) and miR-6879-5p (0.787; 95% CI, 0.706-0.867) and their combination (0.914; 95% CI, 0.865-0.962) were higher than those of the total miRNAs directly isolated from plasma. Moreover, the expression of exosomal miRNAs was stable after treatment with RNase A, prolonged incubation, or repeated freezing and thawing.

CONCLUSIONS

The two plasma exosomal miRNAs (miR-6774-3p and miR-6879-5p) and their combination could serve as new promising biomarkers for the diagnosis of LNM in PTC patients.

摘要

目的

在临床上，甲状腺乳头状癌（PTC）淋巴结转移（LNM）的早期诊断很重要，因为它可以辅助治疗决策，并改善预后。本研究旨在探讨血浆外泌体 miRNA 是否可以作为 PTC 患者 LNM 的潜在诊断标志物。

方法

采用 miRNA 微阵列筛选血浆外泌体 miRNA 谱。在验证集和诊断集中进行定量逆转录聚合酶链反应（qRT-PCR），以选择候选外泌体 miRNA。最后，生成受试者工作特征（ROC）曲线，以评估目标外泌体 miRNA 区分 PTC-N1 患者和 PTC-N0 患者的效率。

结果

在测试集中，共发现 197 个 miRNA 表达差异。基于 qRT-PCR 结果，miR-6774-3p（p<0.001）和 miR-6879-5p（p<0.001）在 PTC-N1 患者中的表达明显高于对照组。血浆外泌体 miR-6774-3p（AUC 值为 0.802；95%CI：0.724-0.879）、miR-6879-5p（AUC 值为 0.787；95%CI：0.706-0.867）及其组合（AUC 值为 0.914；95%CI：0.865-0.962）的 AUC 值均高于直接从血浆中分离的总 miRNA。此外，外泌体 miRNA 的表达在经 RNase A 处理、长时间孵育或反复冻融后仍稳定。