循环 microRNA/isomiRs 作为食管鳞状细胞癌的新型生物标志物。

BACKGROUND: MicroRNA (miR)s are promising diagnostic biomarkers of cancer. Recent next generation sequencer (NGS) studies have found that isoforms of micro RNA (isomiR) circulate in the bloodstream similarly to mature micro RNA (miR). We hypothesized that combination of circulating miR and isomiRs detected by NGS are potentially powerful cancer biomarker. The present study aimed to investigate their application in esophageal cancer. METHODS: Serum samples from patients with esophageal squamous cell carcinoma (ESCC) and age and sex matched healthy control (HC) individuals were investigated for the expression of miR/isomiRs using NGS. Candidate miR/isomiRs which met the criteria in the 1st group (ESCC = 18 and HC = 12) were validated in the 2nd group (ESCC = 30 and HC = 30). A diagnostic panel was generated using miR/isomiRs that were consistently confirmed in the 1st and 2nd groups. Accuracy of the panel was tested then in the 3rd group (ESCC = 18 and HC = 18). Their use was also investigated in 22 paired samples obtained pre- and post-treatment, and in patients with esophageal adenocarcinoma (EAD) and high-grade dysplasia (HGD). RESULTS: Twenty-four miR/isomiRs met the criteria for diagnostic biomarker in the 1st and 2nd group. A multiple regression model selected one mature miR (miR-30a-5p) and two isomiRs (isoform of miR-574-3p and miR-205-5p). The index calculated from the diagnostic panel was significantly higher in ESCC patients than in the HCs (13.3±8.9 vs. 3.1±1.3, p<0.001). The area under the receiver operating characteristics (ROC) curves of the panel index was 0.95. Sensitivity and specificity were 93.8%, and 81% in the 1st and 2nd groups, and 88.9% and 72.3% in the 3rd group, respectively. The panel index was significantly lower in patients with EAD (6.2±4.5) and HGD (4.2±1.7) than in those with ESCC and was significantly decreased at post-treatment compared with pre-treatment (6.2±5.6 vs 11.6±11.5, p = 0.03). CONCLUSION: Our diagnostic panel had high accuracy in the diagnosis of ESCC. MiR/isomiRs detected by NGS could serve as novel biomarkers of ESCC.

背景：微小 RNA（miR）是癌症有前途的诊断生物标志物。最近的下一代测序（NGS）研究发现，微小 RNA（miR）的同种型（isomiR）在血液中循环，类似于成熟的微小 RNA（miR）。我们假设通过 NGS 检测到的循环 miR 和 isomiR 的组合是潜在的强大癌症生物标志物。本研究旨在探讨其在食管癌中的应用。

方法：使用 NGS 检测食管鳞状细胞癌（ESCC）患者和年龄及性别匹配的健康对照（HC）个体的血清样本中 miR/isomiR 的表达。符合第 1 组标准的候选 miR/isomiR（ESCC = 18，HC = 12）在第 2 组（ESCC = 30，HC = 30）中进行验证。使用在第 1 组和第 2 组中一致确认的 miR/isomiR 生成诊断面板。然后在第 3 组（ESCC = 18，HC = 18）中测试该面板的准确性。还在 22 对治疗前后获得的样本以及食管腺癌（EAD）和高级别上皮内瘤变（HGD）患者中研究了它们的用途。

结果：在第 1 组和第 2 组中，有 24 个 miR/isomiR 符合诊断生物标志物标准。多元回归模型选择了一个成熟的 miR（miR-30a-5p）和两个 isomiR（miR-574-3p 和 miR-205-5p 的同种型）。与 HC 相比，ESCC 患者的诊断面板指数显着更高（13.3±8.9 vs. 3.1±1.3，p<0.001）。面板指数的受试者工作特征（ROC）曲线下面积为 0.95。在第 1 组和第 2 组中，敏感性和特异性分别为 93.8%和 81%，在第 3 组中为 88.9%和 72.3%。与 ESCC 患者相比，EAD（6.2±4.5）和 HGD（4.2±1.7）患者的面板指数显着降低，与治疗前相比，治疗后明显降低（6.2±5.6 vs 11.6±11.5，p = 0.03）。

结论：我们的诊断面板在 ESCC 的诊断中具有很高的准确性。通过 NGS 检测到的 miR/isomiR 可作为 ESCC 的新型生物标志物。