Chinese Medical Sciences Journal ›› 2019, Vol. 34 ›› Issue (3): 168-176.doi: 10.24920/003585

• 论著 • 上一篇    下一篇

血清外泌体中CDK6和RHOU mRNA可作为无功能垂体腺瘤侵袭性的标志物

于姗1,王小爽1,曹开灿2,包新杰3,*(),余佳1,*()   

  1. 1 中国医学科学院 北京协和医学院,基础医学研究所,医学分子生物学国家重点实验室RNA与造血调控重点实验室 生物化学与分子生物学系,北京 100005
    2 南方医科大学南方医院胸外科,广州 510515
    3 中国医学科学院 北京协和医学院 北京协和医院神经外科,北京 100730
  • 收稿日期:2019-05-22 出版日期:2019-09-30 发布日期:2019-10-11
  • 通讯作者: 包新杰,余佳 E-mail:xinjieabao@163.com;j-yu@ibms.pumc.edu.cn

Identification of CDK6 and RHOU in Serum Exosome as Biomarkers for the Invasiveness of Non-functioning Pituitary Adenoma

Yu Shan1,Wang Xiaoshuang1,Cao Kaican2,Bao Xinjie3,*(),Yu Jia1,*()   

  1. 1 State Key Laboratory of Medical Molecular Biology & Key Laboratory of RNA and Hematopoietic Regulation & Department of Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & School of Basic Medicine, Peking Union Medical College, Beijing 100005, Chin; 2 Department of Thoracic Surgery, Nanfang Hospital, Guangzhou 510515, China;3 Department of Neurosurgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, Chin;
  • Received:2019-05-22 Published:2019-09-30 Online:2019-10-11
  • Contact: Bao Xinjie,Yu Jia E-mail:xinjieabao@163.com;j-yu@ibms.pumc.edu.cn

摘要:

目的 探索无功能垂体腺瘤(non-functioning pituitary adenomas,NF-PAs)侵袭性的循环分子标志物。

方法 提取侵袭性NF-PAs(invasive NF-PAs,INF-PAs)和非侵袭性NF-PAs(noninvasive NF-PAs,NNF-PAs)患者外周血血清外泌体RNA,采用数字PCR检测候选基因的表达,t检验分析其表达差异。同时利用基因表达值建立受试者工作特征曲线(receiver operating characteristic,ROC),找出阈值,并计算该阈值区分INF-PAs的正确率、灵敏度、特异度和精度,以评估目的基因对侵袭性的诊断效果。

结果 CDK6和RHOU mRNA在INF-PAs中显著上调。ROC曲线显示:CDK6的曲线下面积为0.772(95% CI:0.600-0.943,P=0.005),区分INF-PAs的准确率为77.27%,灵敏度为83.33%,特异度为75.00%,精度为55.56%。RHOU的AUC为0.757(95% CI:0.599-0.915,P=0.007),区分准确率为72.73%,灵敏度为83.33%,特异度为68.75%,精度为50.00%。CDK6和RHOU mRNA在NF-PAs患者血清外泌体中的含量呈正相关(r=0.935, P<0.001)。同时用两个基因的阈值作为预测指标,得到的区分准确率为81.82%,灵敏度为83.33%,特异度为81.25%,精度为62.50%。

结论 血清外泌体中的CDK6和RHOU mRNA分子可作为诊断NF-PA侵袭性的标志物,使用两基因的双阈值诊断可提高准确率、特异度和精度。该结果提示了NF-PAs侵袭性的分子机制,并对侵袭性NF-PAs的早期临床诊断和预后具有重要意义。

关键词: 垂体腺瘤, 侵袭性, 外泌体, 数字PCR, 分子标志物, CDK6, RHOU

Abstract:

Objective To explore circulating biomarkers for screening the invasiveness of non-functioning pituitary adenomas (NF-PAs).

Methods The exosomal RNAs were extracted from serum of patients with invasive NF-PA (INF-PA) or noninvasive NF-PA (NNF-PA). Droplet digital PCR was adapted to detect the mRNA expression of candidate genes related to tumor progression or invasion, such as cyclin dependent kinase 6 (CDK6), ras homolog family member U (RHOU), and spire type actin nucleation factor 2 (SPIRE2). Student’s t-test was used to analyze the statistical difference in the mRNA expression of candidate genes between the two groups. Receiver operating characteristic (ROC) curve was used to establish a model for predicting the invasiveness of NF-PAs. The accuracy, sensitivity, specificity and precision of the model were then obtained to evaluate the diagnostic performance.

Results CDK6 (0.2600±0.0912 vs. 0.1789±0.0628, t=3.431, P=0.0013) and RHOU mRNA expressions (0.2696±0.1118 vs. 0.1788±0.0857, t=2.946, P=0.0052) were upregulated in INF-PAs patients’ serum exosomes as compared to NNF-PAs. For CDK6, the area under the ROC curve (AUC) was 0.772 (95% CI: 0.600-0.943, P=0.005), the accuracy, sensitivity, specificity and precision were 77.27%, 83.33%, 75.00% and 55.56% to predict the invasiveness of NF-PAs. For RHOU, the AUC was 0.757 (95% CI: 0.599-0.915, P=0.007), the accuracy, sensitivity, specificity and precision were 72.73%, 83.33%, 68.75% and 50.00%. In addition, the mRNA levels of CDK6 and RHOU in serum exosomes were significantly positively correlated (r=0.935, P<0.001). After combination of the cut-off scores of the two genes, the accuracy, sensitivity, specificity and precision were 81.82%, 83.33%, 81.25% and 62.50%.

Conclusions CDK6 and RHOU mRNA in serum exosomes can be used as markers for predicting invasiveness of NF-PAs. Combination of the two genes performs better in distinguishing INF-PAs from NNF-PAs. These results indicate CDK6 and RHOU play important roles in the invasiveness of NF-PAs, and the established diagnostic method is valuable for directing the clinical screening and postoperative treatment.

Key words: pituitary adenoma, invasiveness, exosome, droplet digital PCR, biomarkers, RHOU, CDK6

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