胃癌患者血浆环状 RNA 谱分析及其液滴数字 RT-PCR 检测。
Plasma circular RNA profiling of patients with gastric cancer and their droplet digital RT-PCR detection.
机构信息
Department of Biochemistry and Molecular Biology, and Zhejiang Key Laboratory of Pathophysiology, Medical School, Ningbo University, Ningbo, 315211, China.
Department of Gastroenterology, the Affiliated Hospital of Medical School of, Ningbo University, Ningbo, 315020, China.
出版信息
J Mol Med (Berl). 2018 Jan;96(1):85-96. doi: 10.1007/s00109-017-1600-y. Epub 2017 Nov 2.
UNLABELLED
To observe the diagnostic values of circular RNAs (circRNAs), their expression profiles between gastric cancer patients' plasma and healthy controls were first assessed by circRNA microarray. Then, circRNA levels were measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and RT-droplet digital PCR (RT-ddPCR), respectively. A total of 343 differentially expressed circRNAs were found. The top 10 elevated circRNAs in patients were hsa_circ_0088300, hsa_circ_0075825, hsa_circ_0019172, hsa_circ_0000220, hsa_circ_0035277, hsa_circ_0000301, hsa_circ_0000189, hsa_circ_0090080, hsa_circ_0001888, and hsa_circ_0001874. The top 10 reduced circRNAs were hsa_circ_0004771, hsa_circ_0001190, hsa_circ_0061276, hsa_circ_0092337, hsa_circ_0058495, hsa_circ_0061274, hsa_circ_0075829, hsa_circ_0080845, hsa_circ_0001006, and hsa_circ_0003764. In cancer and dysplasia tissues, hsa_circ_0001017 and hsa_circ_0061276 were downregulated. Their levels were significantly associated with distal metastasis. The area under receiver operating characteristic curve in combinative use was 0.966 with 95.5% sensitivity and 95.7% specificity. Patients with low plasma hsa_circ_0001017 or hsa_circ_0061276 had a much shorter overall survival than those with high levels. Patients whose plasma hsa_circ_0001017 or hsa_circ_0061276 levels recovered to normal after operation had a longer disease-free survival. Finally, the in vitro model indicated gastric cancer cells secreting circRNAs into plasma. In conclusion, RT-ddPCR is a potent non-invasive and absolute quantification method for simultaneous detection of multiple circRNAs. Hsa_circ_0001017 and hsa_circ_0061276 are new potential biomarkers for gastric cancer.
KEY MESSAGE
A total of 343 circRNAs are differentially expressed between gastric cancer patients' plasma and healthy controls. Hsa_circ_0001017 and hsa_circ_0061276 are downregulated in gastric cancer tissues. The RT-ddPCR is a potent method for simultaneous detection of multiple circRNAs in plasma. Hsa_circ_0001017 and hsa_circ_0061276 are potential biomarkers for gastric cancer.
目的
通过 circRNA 微阵列首次评估胃癌患者血浆与健康对照者之间环状 RNA（circRNA）的表达谱，观察其诊断价值。然后，分别通过逆转录定量聚合酶链反应（RT-qPCR）和逆转录液滴数字 PCR（RT-ddPCR）测量 circRNA 水平。
方法
共发现 343 个差异表达的 circRNA。患者中前 10 个上调的 circRNA 为 hsa_circ_0088300、hsa_circ_0075825、hsa_circ_0019172、hsa_circ_0000220、hsa_circ_0035277、hsa_circ_0000301、hsa_circ_0000189、hsa_circ_0090080、hsa_circ_0001888 和 hsa_circ_0001874。前 10 个下调的 circRNA 为 hsa_circ_0004771、hsa_circ_0001190、hsa_circ_0061276、hsa_circ_0092337、hsa_circ_0058495、hsa_circ_0061274、hsa_circ_0075829、hsa_circ_0080845、hsa_circ_0001006 和 hsa_circ_0003764。在癌症和发育不良组织中，hsa_circ_0001017 和 hsa_circ_0061276 下调。它们的水平与远端转移显著相关。联合使用的受试者工作特征曲线下面积为 0.966，灵敏度为 95.5%，特异性为 95.7%。血浆 hsa_circ_0001017 或 hsa_circ_0061276 水平低的患者总生存期明显短于 hsa_circ_0001017 或 hsa_circ_0061276 水平高的患者。术后血浆 hsa_circ_0001017 或 hsa_circ_0061276 水平恢复正常的患者无病生存期更长。最后，体外模型表明胃癌细胞将 circRNA 分泌到血浆中。
结论
RT-ddPCR 是一种强大的非侵入性和绝对定量方法，可用于同时检测多种 circRNA。hsa_circ_0001017 和 hsa_circ_0061276 是胃癌的新的潜在生物标志物。
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