外周血 RNA 中 N6-甲基腺苷的水平：胃癌的一种新的预测性生物标志物。

Level of N6-Methyladenosine in Peripheral Blood RNA: A Novel Predictive Biomarker for Gastric Cancer.

机构信息

Department of Clinical Laboratory, Jinling Hospital, Medical School of Nanjing University, Nanjing, China.

Department of Pharmacy, Sun Yat-sen University Cancer Center; State Key Laboratory of Oncology in South China; Collaborative Innovation Center for Cancer Medicine, Guangzhou, China.

出版信息

Clin Chem. 2020 Feb 1;66(2):342-351. doi: 10.1093/clinchem/hvz004.

DOI:10.1093/clinchem/hvz004

PMID:32040577

Abstract

BACKGROUND

Dysregulation of N6-methyladenosine (m6A) is associated with various human diseases including cancer. This study aimed to evaluate the level of m6A as a biomarker for gastric cancer (GC) diagnosis.

METHODS

Peripheral blood samples were collected from 100 GC patients, 30 benign gastric disease (BGD) patients, and 75 healthy controls (HCs). Levels of m6A in total RNA and expression of m6A-related proteins were analyzed.

RESULTS

The m6A levels in peripheral blood RNA were significantly increased in the GC group compared with those in the BGD or HC groups; moreover, levels increased with the progression and metastasis of GC and decreased in GC patients after surgery. The area under the curve (AUC) for m6A in the GC group was 0.929 (95% CI, 0.88-0.96), which is markedly greater than the AUCs for carcinoembryonic antigen (CEA; 0.694) and carbohydrate antigen 199 (CA199; 0.603). The combination of CEA and CA199 with m6A improved the AUC to 0.955 (95% CI, 0.91-0.98). The expressions of m6A demethylases ALKBH5 and FTO were significantly downregulated in the GC group compared with the HC group. Coculture with GC cells increased the m6A of RNA in promyelocytic (HL-60) and monocytic (THP-1) leukemia cells and nontumorigenic human peripheral blood B lymphocyte cells (PENG-EBV). Furthermore, a xenograft model enhanced the m6A in peripheral blood RNA of mice. Accordingly, expressions of ALKBH5 and FTO were decreased both in vitro and in vivo.

CONCLUSIONS

Level of m6A in peripheral blood RNA is a promising noninvasive diagnostic biomarker for GC patients.

摘要

背景

N6-甲基腺苷（m6A）的失调与包括癌症在内的各种人类疾病有关。本研究旨在评估 m6A 作为胃癌（GC）诊断生物标志物的水平。

方法

收集 100 例 GC 患者、30 例良性胃部疾病（BGD）患者和 75 例健康对照（HC）的外周血样本。分析总 RNA 中的 m6A 水平和 m6A 相关蛋白的表达。

结果

GC 组外周血 RNA 中的 m6A 水平明显高于 BGD 组或 HC 组；此外，水平随 GC 的进展和转移而增加，并在 GC 患者手术后降低。GC 组 m6A 的曲线下面积（AUC）为 0.929（95%CI，0.88-0.96），明显大于癌胚抗原（CEA；0.694）和糖类抗原 199（CA199；0.603）的 AUC。CEA 和 CA199 与 m6A 的联合使用将 AUC 提高到 0.955（95%CI，0.91-0.98）。与 HC 组相比，GC 组的 m6A 去甲基酶 ALKBH5 和 FTO 的表达明显下调。与 GC 细胞共培养可增加 HL-60 早幼粒细胞白血病细胞和 THP-1 单核细胞白血病细胞以及非致瘤性人外周血 B 淋巴细胞（PENG-EBV）中 RNA 的 m6A。此外，异种移植模型增强了小鼠外周血 RNA 中的 m6A。因此，ALKBH5 和 FTO 的表达在体外和体内均降低。