Expression Profile of Serum-related Exosomal miRNAs from Parathyroid Tumor

Objective: The expression pattern of exosomal miRNAs derived from parathyroid tumor is still unknown. In the present work, we aimed to examine the differences on microRNA (miRNA) expression, present in serum exosomes, by comparing parathyroid carcinoma (PC) and parathyroid adenoma (PA). Methods: MiRNA expression profile of serum exosomes, derived from 4 PC patients and 4 PA patients, were analyzed by next-generation sequencing technology. The differential expressions of target miRNAs were further verified in both serum exosomes and tissues of PC/PA patients by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Lastly, receiver operating characteristic (ROC) curves were plotted to investigate the efficiency of target exosomal miRNAs in distinguishing PC patients from PA controls. Results: Multiple differentially expressed miRNAs of serum exosomes were screened out by sequencing. Based on this screening, hsa-miR-146b-5p (p=0.0846), hsa-miR-27a-5p (p=0.0412), hsa-miR-93-5p (p=0.73), hsa-miR-381-3p (p=0.1239) and hsa-miR-134-5p (p=0.0694) were up-regulated in the serum exosomes of PC patients. These results were validated by qPCR, where the trend on differential miRNA expression was consistent with the sequencing results. Specifically, the expression of exosomal hsa-miR-27a-5p was able to clearly distinguish PC patients from PA controls, and related analysis indicated that the area under the ROC curve was 0.8594 (p=0.0157). Conclusion: Here we present, for the first time, the miRNA expression profile of serum exosomes derived from PC patients. Based on this result, we presently suggest that the exosomal hsa-miR-27a-5p may serve as a putative tumor marker for preoperative identification of PC and PA subjects. A total of 25 PHPT patients, including 12 PC and 13 PA patients, were included in this study. 4 cases of specimens were randomly selected from the PC (2 males and 2 females) and PA (1 male and 3 females) groups for sequencing. The remaining specimens in the PC and PA groups, together with 8 NC, were utilized for verification and analysis.

研究目的：甲状旁腺肿瘤来源的外泌体microRNA（miRNA）的表达模式至今尚未明确。本研究旨在通过比较甲状旁腺癌（parathyroid carcinoma, PC）与甲状旁腺腺瘤（parathyroid adenoma, PA）患者血清外泌体中的miRNA表达差异，探究相关表达谱变化。
研究方法：本研究共纳入25例原发性甲状旁腺功能亢进症（PHPT）患者，其中12例PC患者、13例PA患者。从PC组（2男2女）与PA组（1男3女）中随机各选取4例标本，采用下一代测序技术分析其血清外泌体的miRNA表达谱。随后，采用定量反转录聚合酶链反应（qRT-PCR），在PC/PA患者的血清外泌体及组织中验证靶miRNA的差异表达情况。最后，绘制受试者工作特征（ROC）曲线，评估靶外泌体miRNA区分PC患者与PA对照的效能。剩余的PC组、PA组标本，以及8例正常对照（normal control, NC）标本，则用于后续的验证与分析。
研究结果：通过测序筛选出多个血清外泌体差异表达miRNA。基于筛选结果，PC患者血清外泌体中hsa-miR-146b-5p（p=0.0846）、hsa-miR-27a-5p（p=0.0412）、hsa-miR-93-5p（p=0.73）、hsa-miR-381-3p（p=0.1239）及hsa-miR-134-5p（p=0.0694）呈上调表达。上述结果经qPCR验证，miRNA差异表达趋势与测序结果一致。其中，外泌体hsa-miR-27a-5p的表达可有效区分PC患者与PA对照，相关分析显示其ROC曲线下面积为0.8594（p=0.0157）。
研究结论：本研究首次报道了PC患者来源的血清外泌体miRNA表达谱。基于上述结果，本研究提示外泌体hsa-miR-27a-5p可作为潜在的肿瘤标志物，用于术前鉴别PC与PA患者。