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中华肝脏外科手术学电子杂志 ›› 2025, Vol. 14 ›› Issue (03) : 463 -470. doi: 10.3877/cma.j.issn.2095-3232.2025.03.020

基础研究

CEBPZOS通过调控肿瘤增殖与迁移促进肝癌进展的机制研究
张铭燊1, 胡永威1, 陈德盛1, 俞浩远1, 梁智星1, 陈玉涛1, 叶林森1, 李华1, 杨扬1,()   
  1. 1. 510630 广州,中山大学附属第三医院肝脏外科暨肝移植中心 广东省器官移植研究中心 广东省肝脏疾病研究重点实验室
  • 收稿日期:2025-02-05 出版日期:2025-06-10
  • 通信作者: 杨扬
  • 基金资助:
    国家自然科学基金面上项目(81972286)广东省自然科学基金面上项目(2023A1515010322)国家资助博士后研究人员计划C档资助(GZC20242089)广州市自然科学基金重点项目(02A004999000186)

Study on mechanism of CEBPZOS in promoting progression of hepatocellular carcinoma by regulating tumor proliferation and migration

Mingshen Zhang1, Yongwei Hu1, Desheng Chen1, Haoyuan Yu1, Zhixing Liang1, Yutao Chen1, Linsen Ye1, Hua Li1, Yang Yang1,()   

  1. 1. Department of Liver Surgery & Liver Transplantation Center,the Third Affiliated Hospital of Sun Yat-sen University,Organ Transplantation Research Center of Guangdong Province,Guangdong Provincial Key Laboratory of Liver Diseases Research,Guangzhou 510630,China
  • Received:2025-02-05 Published:2025-06-10
  • Corresponding author: Yang Yang
引用本文:

张铭燊, 胡永威, 陈德盛, 俞浩远, 梁智星, 陈玉涛, 叶林森, 李华, 杨扬. CEBPZOS通过调控肿瘤增殖与迁移促进肝癌进展的机制研究[J/OL]. 中华肝脏外科手术学电子杂志, 2025, 14(03): 463-470.

Mingshen Zhang, Yongwei Hu, Desheng Chen, Haoyuan Yu, Zhixing Liang, Yutao Chen, Linsen Ye, Hua Li, Yang Yang. Study on mechanism of CEBPZOS in promoting progression of hepatocellular carcinoma by regulating tumor proliferation and migration[J/OL]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2025, 14(03): 463-470.

目的

探讨CCAAT增强子结合蛋白ζ反义链(CEBPZOS)在肝癌组织中的表达及促进肝细胞癌(肝癌)发展的机制。

方法

基于TCGA和GTEx数据库分析CEBPZOS表达与临床分期、预后的相关性;构建CEBPZOS敲低/过表达肝癌细胞系Hepa1-6,并通过CCK-8实验、克隆形成实验、Transwell迁移实验、细胞划痕实验评估CEBPZOS敲低/过表达对Hepa1-6细胞增殖迁移的影响;构建CEBPZOS敲低/过表达肝癌细胞系Hepa1-6-luc及皮下肝癌小鼠模型,采用小动物体内成像动态监测肿瘤辐射强度变化,同时监测小鼠体质量变化。组间实验数据比较采用单因素方差分析,生存分析采用Kaplan-Meier法和Log-rank检验。

结果

基于TCGA数据库及GTEx数据库分析显示,肝癌组织中CEBPZOS的mRNA表达水平明显高于癌旁正常组织(P<0.05);随着肿瘤进展(Ⅰ~Ⅲ期),CEBPZOS mRNA表达呈递增趋势(F=6.41,P<0.05)。CEBPZOS高表达组总体生存率及无病生存率明显低于低表达组(HR=2.00,1.70;P<0.05)。细胞增殖实验及克隆形成实验结果显示,CEBPZOS敲低后Hepa1-6细胞的增殖能力明显下降, 而CEBPZOS过表达Hepa1-6细胞增殖能力增强(F=148.60,223.80,P<0.05);细胞划痕实验和Transwell迁移实验结果显示,CEBPZOS敲低Hepa1-6细胞的迁移能力下降,CEBPZOS过表达Hepa1-6细胞迁移能力增强(F=387.50,80.97;P<0.05)。动物实验显示,CEBPZOS过表达组小鼠肿瘤辐射强度随时间增强(F=142.80,P<0.05);CEBPZOS过表达组的5、9、14 d肿瘤辐射强度最高,CEBPZOS敲低组肿瘤辐射强度最低(F=5.24,52.77,111.00;P<0.05);CEBPZOS过表达组10、12、14 d的体质量增加明显(F=10.27,28.70,29.25;P<0.05)。

结论

CEBPZOS在肝癌中高表达,CEBPZOS高表达与肿瘤快速进展、较晚的TNM分期及患者不良预后密切相关。CEBPZOS可过促进肿瘤细胞增殖、迁移促进肝癌进展,是潜在预后标志物和治疗靶点。

Objective

To investigate the expression level of CCAAT enhancer binding protein ζ antisense strand (CEBPZOS) in hepatocellular carcinoma (HCC) tissues and the mechanism of CEBPZOS in promoting the progression of HCC.

Methods

Based on TCGA and GTEx databases, the correlation between the expression of CEBPZOS and clinical stages and prognosis was analyzed. Hepa1-6 cell lines with CEBPZOS knockdown/overexpression were constructed. The effect of CEBPZOS knockdown/overexpression on the proliferation and migration of Hepa1-6 cells was assessed by CCK-8 assay, colony formation assay,Transwell migration chamber and scratch assay, respectively. Hepa1-6-luc cell lines with CEBPZOS knockdown/overexpression and mouse models with subcutaneous HCC were constructed. The changes of tumor radiation intensity were dynamically monitored by in vivo imaging of small animals. Meantime,the changes in body weight of mice were also recorded. The experimental data between two groups were compared by one-way ANOVA. Survival analysis was performed by Kaplan-Meier method and Log-rank test.

Results

Based on TCGA and GTEx database analyses, the expression level of CEBPZOS mRNA in liver cancer tissues was significantly higher than that in normal tissues adjacent to liver cancer (P<0.05). With the progression of tumors (stageⅠ-Ⅲ), the expression level of CEBPZOS mRNA showed an increasing trend(F=6.41, P<0.05). The overall survival and disease-free survival rates in the CEBPZOS overexpression group were significantly lower than those in the CEBPZOS knockdown group (HR=2.00,1.70; both P<0.05). CCK-8 and colony formation assays in vitro showed that the proliferation ability of Hepa1-6 cells was significantly decreased after CEBPZOS knockdown, whereas that of Hepa1-6 cells with CEBPZOS overexpression was enhanced (F=148.60, 223.80, both P<0.05). Scratch test and Transwell migration chamber revealed that the migration ability of Hepa1-6 cells was decreased after CEBPZOS knockdown, while that of Hepa1-6 cells was increased after CEBPZOS overexpression (F=387.50,80.97; both P<0.05). Animal experiments showed that the tumor radiation intensity of mice in the CEBPZOS overexpression group was significantly enhanced over time (F=142.80, P<0.05). On the 5th, 9th and 14th d, the tumor radiation intensity was the highest in the CEBPZOS overexpression group, while that in the CEBPZOS knockdown group was the lowest (F=5.24,52.77, 111.00; all P<0.05). The body weight of mice in the CEBPZOS overexpression group was significantly increased on the 10th, 12th and 14th d (F=10.27, 28.70, 29.25; all P<0.05).

Conclusions

CEBPZOS is highly expressed in HCC, which is closely associated with rapid progression of tumors, high TNM stage and poor prognosis. CEBPZOS can promote the progression of HCC by accelerating the proliferation and migration of tumor cells, which is a potential prognostic biomarker and therapeutic target.

图1 CEBPZOS在肝癌中表达及病理学特征
图2 CEBPZOS表达与肝癌患者生存的相关性
图3 CEBPZOS表达对肝癌细胞增殖、迁移能力的影响
图4 CEBPZOS表达水平对皮下肝癌小鼠模型肿瘤生长的影响
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