Institute of Organ Transplantation, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Cancer Biomark. 2021;32(2):137-146. doi: 10.3233/CBM-203169.
Long non-coding RNA-growth arrest specific transcript 5 (lncRNA-GAS5) plays a suppressive role in activated hepatic stellate cells (HSCs). LncRNAs could circulate in the blood in a cell-free form and serve as promising biomarkers for various human diseases. Herein, we investigated the feasibility of using serum GAS5 as a biomarker for liver fibrosis in chronic hepatitis B (CHB) patients and whether promoter methylation was responsible for GAS5 down-regulation.
Serum GAS5 levels were quantified using quantitative real-time PCR in CHB patients and healthy controls. GAS5 promoter methylation was examined in LX-2 cells and cirrhotic tissues.
Compared with the sera from healthy controls, lower GAS5 levels were found in the sera from CHB patients. Receiver operating characteristic curve analysis indicated that serum GAS5 had a significant diagnostic value for liver fibrosis in CHB patients. Serum GAS5 negatively correlated with HAI scores as well as ALT values in CHB patients. GAS5 was additionally reduced in cirrhotic tissues, associated with its hypermethylation promoter. In LX-2 cells, transforming growth factor-β1 treatment led to a reduction in GAS5 expression and an increase in promoter methylation. Hypermethylation of GAS5 was blocked down by DNA methyltransferase (DNMT) inhibitor and restored GAS5 inhibited HSC activation including proliferation and collagen production. Further studies confirmed that GAS5 methylation was mediated by DNMT1.
We demonstrate that epigenetically-regulated serum GAS5 could serve as a potential biomarker in CHB patients. Loss of GAS5 is associated with DNMT1-mediated promoter methylation.
长链非编码 RNA-生长停滞特异性转录物 5（lncRNA-GAS5）在激活的肝星状细胞（HSCs）中发挥抑制作用。lncRNAs 可以以无细胞形式在血液中循环，并作为各种人类疾病的有前途的生物标志物。在此，我们研究了使用血清 GAS5 作为慢性乙型肝炎（CHB）患者肝纤维化的生物标志物的可行性，以及启动子甲基化是否负责 GAS5 下调。
使用定量实时 PCR 定量检测 CHB 患者和健康对照者的血清 GAS5 水平。在 LX-2 细胞和肝硬化组织中检查 GAS5 启动子甲基化。
与健康对照者的血清相比，CHB 患者的血清中 GAS5 水平较低。受试者工作特征曲线分析表明，血清 GAS5 对 CHB 患者的肝纤维化具有显著的诊断价值。血清 GAS5 与 CHB 患者的 HAI 评分和 ALT 值呈负相关。GAS5 在肝硬化组织中进一步减少，与其启动子高甲基化相关。在 LX-2 细胞中，转化生长因子-β1 处理导致 GAS5 表达减少和启动子甲基化增加。DNA 甲基转移酶（DNMT）抑制剂可阻断 GAS5 的高甲基化，并恢复 GAS5 抑制 HSC 激活，包括增殖和胶原产生。进一步的研究证实 GAS5 甲基化是由 DNMT1 介导的。
我们证明了受表观遗传调控的血清 GAS5 可作为 CHB 患者的潜在生物标志物。GAS5 的丢失与 DNMT1 介导的启动子甲基化有关。
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