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中华实验和临床感染病杂志(电子版) ›› 2026, Vol. 20 ›› Issue (02) : 106 -113. doi: 10.3877/cma.j.issn.1674-1358.2026.02.006

论著

人类Runt相关转录因子、E-钙黏蛋白和Ras相关区域家族1A基因甲基化对丙型肝炎病毒相关肝细胞癌的诊断价值
蔡轶伦1, 陈燕2, 董金玲2,†()   
  1. 1 313000 湖州市，湖州师范大学医学院（护理学院）
    2 313000 湖州市，湖州师范学院附属第一医院（湖州市第一人民医院）感染科
  • 收稿日期:2025-09-03 出版日期:2026-04-15
  • 通信作者: 董金玲
  • 基金资助:
    湖州市科技计划项(2021GY19)

Diagnostic value of human Runt-related transcription factor 3, E-cadherin, and Ras association domain family 1A gene methylation in hepatitis C virus-associated hepatocellular carcinoma

Yilun Cai1, Yan Chen2, Jinling Dong2,†()   

  1. 1 School of Medicine (School of Nursing), Huzhou Normal University, Huzhou 313000, China
    2 Infectious Disease Department, Huzhou First People’s Hospital, the First Affiliated Hospital of Huzhou University, Huzhou 313000, China
  • Received:2025-09-03 Published:2026-04-15
  • Corresponding author: Jinling Dong
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引用本文：

蔡轶伦, 陈燕, 董金玲. 人类Runt相关转录因子、E-钙黏蛋白和Ras相关区域家族1A基因甲基化对丙型肝炎病毒相关肝细胞癌的诊断价值[J/OL]. 中华实验和临床感染病杂志(电子版), 2026, 20(02): 106-113.

Yilun Cai, Yan Chen, Jinling Dong. Diagnostic value of human Runt-related transcription factor 3, E-cadherin, and Ras association domain family 1A gene methylation in hepatitis C virus-associated hepatocellular carcinoma[J/OL]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2026, 20(02): 106-113.

目的

探讨人类Runt相关转录因子（RUNX3）、E-钙黏蛋白（E-cadherin）和Ras相关区域家族1A（RASSF1A）基因甲基化对丙型肝炎病毒（HCV）相关肝细胞癌（HCC）的诊断价值。

方法

回顾性分析2022年6月至2024年2月湖州市第一人民医院收治的30例HCV相关HCC患者（HCC组）、30例HCV相关肝硬化患者（肝硬化组）、30例单纯HCV感染者（HCV组）和30例健康体检者（健康对照组）的临床资料，比较4组研究对象的一般资料、甲胎蛋白（AFP）（筛查HCC的传统指标）以及RUNX3、E-Cadherin和RASSF1A基因甲基化水平；比较各组研究对象以上3种基因甲基化水平对应的AFP水平；采用受试者工作特征（ROC）曲线分析3种基因甲基化与AFP分别及联合检测对HCV相关HCC的诊断效能；应用多因素Logistic回归分析3种基因甲基化和AFP水平是否为发生HCV相关HCC的影响因素。

结果

各组研究对象TBil（F=339.823、P＜0.001）、ALT（F=681.714、P＜0.001）、AFP（F=1 488.962、P＜0.001）水平，RUNX3甲基化（F=92.207、P＜0.001）、E-cadherin甲基化（F=39.316、P＜0.001）和RASSF1A甲基化（F=21.705、P＜0.001）水平差异均有统计学意义。RUNX3、E-Cadherin和RASSF1A基因高甲基化与AFP联合诊断HCV相关HCC的ROC曲线下面积（AUC）为0.916，灵敏度和特异度分别为82.50%和55.00%。HCC组患者RUNX3甲基化低水平（＜0.23%）和高水平（≥0.23%），E-cadherin甲基化低水平（＜0.14%）和高水平（≥0.14%）以及RASSF1A甲基化低水平（＜0.41%）和高水平（≥0.41%）对应的AFP水平差异均有统计学意义（t=3.304、P=0.002，t=4.495、P＜0.001，t=3.158、P=0.003）。多因素Logistic回归分析显示，RUNX3基因甲基化水平（OR=8.306、95%CI：1.862～37.058、P=0.006）、RASSF1A基因甲基化水平（OR=7.205、95%CI：2.080～25.268、P=0.002）和AFP水平（OR=8.793、95%CI：1.725～44.825、P=0.009）均为HCV感染者发生HCC的危险因素，E-cadherin甲基化水平为HCV感染者发生HCC的保护因素（OR=0.457、95%CI：0.122～0.897、P=0.025）。

结论

血浆RASSF1A、E-cadherin和RUNX3甲基化可作为HCV相关HCC有效诊断指标，并可能在预测肝硬化进展为HCC发挥重要作用。

关键词: 肝炎病毒，丙型, 原发性肝细胞癌, 肝硬化, 人类Runt相关转录因子, E-钙黏蛋白, Ras相关区域家族1A, 甲基化
Objective

To investigate the diagnostic value of plasma methylation levels of human Runt-related transcription factor 3 (RUNX3), E-cadherin and Ras-association domain family 1A (RASSF1A) genes in hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC).

Methods

Clinical data of 30 patients with HCV-related HCC (HCC group), 30 patients with HCV-related liver cirrhosis (liver cirrhosis group), 30 patients with simple HCV infection (HCV group) and 30 healthy individuals undergoing physical examination (healthy control group) admitted to Huzhou First People’s Hospital from June 2022 to February 2024 were analyzed, retrospectively. General data, alpha-fetoprotein (AFP) which was a traditional indicator for HCC screening, and methylation levels of gene RUNX3, E-cadherin and RASSF1A were compared among the four groups, respectively. The levels of AFP corresponding to high and low methylation levels of the three genes were compared in each group. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic efficacy of the three gene methylations and AFP alone or in combination for HCV-related HCC. Multivariate Logistic regression was applied to analyze whether the three gene methylations and AFP level were influencing factors for HCV-related HCC.

Results

There were significant differences in total bilirubin (TBil) (F=339.823, P<0.001), alanine aminotransferase (ALT) (F=681.714, P<0.001), AFP (F=1488.962, P<0.001), and methylation levels of RUNX3 (F=92.207, P<0.001), E-cadherin (F=39.316, P<0.001) and RASSF1A (F=21.705, P<0.001) among the four groups. The area under the ROC curve (AUC) of the combined diagnosis of HCV-related HCC by hypermethylation of RUNX3, E-cadherin and RASSF1A genes plus AFP was 0.916, with the sensitivity of 82.50% and specificity of 55.00%. In the HCC group, AFP levels corresponding to low (<0.23%) and high (≥0.23%) RUNX3 methylation levels, low (<0.14%) and high (≥0.14%) E-cadherin methylation levels, as well as low (<0.41%) and high (≥0.41%) RASSF1A methylation levels all showed significant differences (t=3.304, P=0.002; t=4.495, P<0.001; t=3.158, P=0.003). Multivariate Logistic regression analysis showed that RUNX3 methylation (OR=8.306, 95%CI: 1.862-37.058, P=0.006), RASSF1A methylation (OR=7.205, 95%CI: 2.080-25.268, P=0.002), and AFP level (OR=8.793, 95%CI: 1.725-44.825, P=0.009) were all risk factors for HCC in HCV-infected patients, while E-cadherin methylation was a protective factor (OR=0.457, 95%CI: 0.122-0.897, P=0.025).

Conclusions

Plasma methylation levels of RASSF1A, E-cadherin and RUNX3 gene can serve as effective diagnostic indicators for HCV-related HCC and may play important roles in predicting the progression of liver cirrhosis to HCC.

Key words: Hepatitis C virus, Hepatocellular carcinoma, Liver cirrhosis, Human Runt-related transcription factor 3, E-cadherin, Ras association domain family 1A, Methylation
表1 甲基化特异性PCR反应体系
反应体系 体积（µl）
TaKaRa Ex HS Taq（5 U/µl） 0.125
10× Ex Taq Buffer（Mg2+plus） 2.5
dNTP（各2.5 µmol/L） 2
M/U上游引物（10 µmol/L） 0.5
M/U下游引物（10 µmol/L） 0.5
模板DNA 2
灭菌蒸馏水 17.375
合计 25
表2 甲基化特异性PCR引物设计
基因 引物序列（5'→3'）
E-cadherin-M
正向 TGTAGTTACGTATTTATTTTTAGTGGCGTC
反向 CGAATACGATCGAATCGAACCG
E-cadherin-U
正向 TGGTTGTAGTTATGTATTTATTTTTAGTGGTGTT
反向 ACACCAAATACAATCAAATCAAACCAAA
RASSF1A-M
正向 GTGTTAACGCGTTGCGTATC
反向 AACCCCGCGAACTAAAAACGA
RASSF1A-U
正向 TTTGGTTGGAGTGTGTTAATGTG
反向 CAAACCCCACAAACTAAAAACAA
RUNX3-M
正向 TTACGAGGGGCGGTCGTACGCGGG
反向 AAAACGACCGACGCGAACGCCTCC
RUNX3-U
正向 TTATGAGGGGTGGTTGTATGTGGG
反向 AAAACAACCAACACAAACACCTCC
表3 各组研究对象的一般资料[例（%）]
一般资料 健康对照组（30例） HCV组（30例） 肝硬化组（30例） HCC组（30例） 统计量 P值
性别 [例（%）] χ2=0.779 0.859a
男 19（63.30） 21（70.00） 20（66.70） 22（73.30）
女 11（36.70） 9（30.00） 10（33.30） 8（26.70）
年龄（
±s，岁）		 55.23 ± 10.12 52.96 ± 12.63 53.88 ± 11.01 54.30 ± 12.63 F=0.262 0.853
腹腔积液 [例（%）] — — 3（7.50） 5（12.50） χ2=0.556 0.456a
TBil（
±s，μmol/L）		 13.75 ± 4.36 14.44 ± 3.12 26.36 ± 2.36 36.74 ± 4.69 F=339.823 ＜0.001
ALT（
±s，U/L）		 5.74 ± 1.20 14.38 ± 2.00 63.85 ± 10.74 104.63 ± 11.63 F=681.714 ＜0.001
AFP（
±s，ng/ml）		 21.67 ± 4.79 23.60 ± 4.68 181.72 ± 29.81 206.11 ± 18.13 F=1 488.962 ＜0.001
表4 各组研究对象RUNX3、E-Cadherin和RASSF1A基因甲基化水平（
±s，%）
组别 例数 RUNX3甲基化 E-Cadherin甲基化 RASSF1A甲基化
健康对照组 30 0.09 ± 0.02 0.05 ± 0.01 0.24 ± 0.10
HCV组 30 0.13 ± 0.03 0.06 ± 0.02 0.28 ± 0.11
肝硬化组 30 0.17 ± 0.03 0.08 ± 0.03 0.35 ± 0.11
HCC组 30 0.24 ± 0.07 0.12 ± 0.05 0.44 ± 0.15
F值 92.207 39.316 21.705
P值 ＜0.001 ＜0.001 ＜0.001
图1 RUNX3、E-Cadherin和RASSF1A基因甲基化与AFP水平诊断HCV相关HCC的ROC曲线
表5 RUNX3、E-Cadherin和RASSF1A基因甲基化与AFP水平对HCV相关HCC的诊断效能
指标 AUC 截断值 95%CI 敏感度（%） 特异度（%）
RUNX3 0.797 0.23% 0.688～0.905 62.50 98.50
E-Cadherin 0.719 0.14% 0.607～0.831 37.50 97.50
RASSF1A 0.666 0.41% 0.544～0.788 67.50 70.00
AFP 0.766 187.88 ng/ml 0.660～0.873 72.00 62.50
联合检测 0.916 — 0.858～0.974 82.50 55.00
表6 各组研究对象RUNX3、E-Cadherin和RASSF1A基因甲基化水平对应的AFP水平（
±s，ng/ml）
基因甲基化 健康对照组 HCV组 肝硬化组 HCC组
例数 AFP 例数 AFP 例数 AFP 例数 AFP
RUNX3
≥0.23% 0 — 0 — 0 — 19 213.40 ± 11.75
＜0.23% 30 5.74 ± 1.20 30 14.38 ± 2.00 40 181.72 ± 29.81 11 193.92 ± 20.87
t值 — 3.304
P值 — 0.002
E-cadherin
≥0.14% 0 — 0 — 1 174.4 11 219.82 ± 13.94
＜0.14% 30 5.74 ± 1.20 30 14.38 ± 2.00 29 181.90 ± 27.50 19 197.95 ± 15.43
t值 — — — 4.495
P值 — — — ＜0.001
RASSF1A
≥0.41% 1 6.14 2 15.23 ± 1.20 9 197.46 ± 16.74 20 211.05 ± 17.55
＜0.41% 29 5.73 ± 1.51 28 14.32 ± 2.08 21 174.96 ± 25.68 10 196.05 ± 12.46
t值 — 0.742 2.781 3.158
P值 — 0.463 0.008 0.003
表7 发生HCV相关HCC影响因素的多因素Logistic回归分析赋值
变量 赋值
组别 肝硬化组=0；HCC组=1
RUNX3 ＜0.23%=0；≥0.23%=1
E-Cadherin ＜0.14%=0；≥0.14%=1
RASSF1A ＜0.41%=0；≥0.41%=1
AFP ＜187.88 ng/ml=0；≥187.88 ng/ml=1
表8 RUNX3、E-Cadherin、RASSF1A甲基化和AFP水平是否为发生HCV相关HCC影响因素的多因素Logistic回归分析
变量 β值 S.E.值 Wald χ2值 OR值 95%CI P值
RUNX3 2.117 0.763 7.698 8.306 1.862～37.058 0.006
E-Cadherin 0.899 0.400 5.051 0.457 0.122～0.897 0.025
RASSF1A 1.981 0.637 9.671 7.205 2.080～25.268 0.002
AFP 2.174 0.831 6.844 8.793 1.725～44.825 0.009
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