Measurement of Cyclin D2 (CCND2) Gene Promoter Methylation in Plasma and Peripheral Blood Mononuclear Cells and Alpha-Fetoprotein Levels in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma
Yu Qian 1 , Jing-Wen Wang 1 , Yu Fang 1 , Xiao-Dong Yuan 1 , Yu-Chen Fan 1 2 , Shuai Gao 1 2 , Kai Wang 1 2
Affiliations
Affiliations
1
Department of Hepatology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China (mainland).
2
Institute of Hepatology, Shandong University, Jinan, Shandong, China (mainland).
PMID: 33320844 DOI: 10.12659/MSM.927444
Abstract
BACKGROUND Alpha-fetoprotein (AFP) is widely used to screen for hepatocellular carcinoma (HCC). However, the use of this biomarker has been challenged due to its low sensitivity and high rate of false negatives. In this study, we evaluated the diagnostic capability of cyclin D2 (CCND2) promoter methylation in patients with HCC related to hepatitis B virus (HBV). MATERIAL AND METHODS Using methylation-specific PCR and quantitative real-time PCR, we measured methylation status and mRNA levels of CCND2 in plasma and peripheral blood mononuclear cells (PBMCs) from 275 subjects: 75 patients with chronic hepatitis B (CHB), 47 with liver cirrhosis (LC), 118 with HCC, and 35 healthy controls (HCs). RESULTS The methylation rate of the CCND2 promoter was significantly higher in HCC patients than in patients without HCC (P<0.001). Furthermore, advanced HCC (TNM III/IV) was associated with a significantly higher frequency of CCND2 methylation and lower CCND2 mRNA levels than early-stage disease (TNM I/II; P<0.05). Combined measurement of CCND2 methylation and AFP yielded significantly higher sensitivity and area under the curve (AUC) than AFP alone in distinguishing patients with HCC from subjects with LC and CHB (P<0.001). CONCLUSIONS CCND2 methylation may be useful for predicting HCC progression. In addition, combined measurement of CCND2 methylation and AFP could serve as a non-invasive diagnostic marker for patients with HBV-related HCC. 作者: StephenW 时间: 2020-12-16 14:52
背景技术甲胎蛋白（AFP）被广泛用于筛查肝细胞癌（HCC）。但是，由于该生物标记物的低灵敏度和高假阴性率，因此其使用受到了挑战。在这项研究中，我们评估了细胞周期蛋白D2（CCND2）启动子甲基化在与乙型肝炎病毒（HBV）相关的HCC患者中的诊断能力。材料和方法使用甲基化特异性PCR和定量实时PCR，我们测量了275名受试者的血浆和外周血单核细胞（PBMC）中CCND2的甲基化状态和mRNA水平：75例慢性乙型肝炎（CHB），47例肝硬化（LC），118例HCC和35例健康对照（HCs）。结果HCC患者CCND2启动子的甲基化率显着高于无HCC患者（P <0.001）。此外，与早期疾病相比，晚期肝癌（TNM III / IV）与CCND2甲基化的频率明显升高和CCND2 mRNA的水平降低（TNM I / II； P <0.05）。与单独使用AFP相比，CCND2甲基化和AFP的联合测量产生的灵敏度和曲线下面积（AUC）明显更高，从而可以将HCC患者与LC和CHB患者区分开来（P <0.001）。结论CCND2甲基化可能有助于预测HCC进程。此外，CCND2甲基化和AFP的联合测量可以作为HBV相关性HCC患者的非侵入性诊断标记。