ADH1B对肝细胞癌合并脉管侵犯患者的预后价值及肝癌生物学行为的影响  

Prognostic value of ADH1B in patients with hepatocellular carcinoma with vascular invasion and its effect on the biological behavior of liver cancer

作　　者：覃水玲 徐静轩 韦浩文 马良[1] 齐鲁楠[1,2] QIN Shuiling;XU Jingxuan;WEI Haowen;MA Liang;QI Lunan(Department of Hepatobiliary Surgery,Guangxi Medical University Cancer Hospital,Nanning 530021,China;Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor,Ministry of Education,Nanning 530021,China)

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,南宁530021 [2]广西区域性高发肿瘤早期防治研究重点实验室,南宁530021

出　　处：《中国癌症防治杂志》2023年第4期383-390,共8页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：国家自然科学基金面上项目(81972306)。

摘　　要：目的探讨乙醇脱氢酶1B(alcohol dehydrogenase 1B,ADH1B)分子在合并脉管侵犯的肝细胞癌(hepatocellular carcinoma with vascular invasion,VI-HCC)患者中的临床价值及对肝癌细胞生物学功能的影响。方法对VI-HCC组织进行RNA测序及GO、KEGG、GSEA、WGCNA等分析。采用免疫组织化学法检测组织中的ADH1B蛋白表达水平。采用Kaplan-Meier法绘制生存曲线并运用log-rank检验分析不同ADH1B蛋白表达组患者的生存差异,单因素及多因素Cox回归模型分析影响VI-HCC患者术后预后的独立危险因素,Spearman检验分析ADH1B与AFP、Ki67的相关性。用过表达ADH1B及其阴性对照的慢病毒感染肝癌Huh7细胞以构建ADH1B过表达的稳转株,采用qRT-PCR法检测细胞中ADH1B mRNA的表达水平。采用EDU、平板克隆形成实验检测细胞的增殖能力,Transwell侵袭、迁移实验及划痕实验检测细胞的侵袭、迁移能力,流式细胞术检测细胞的凋亡能力。结果RNA测序结果发现VI-HCC根治术后预后不良的患者原发瘤呈代谢重编程特征。免疫组织化学结果显示VI-HCC的癌组织中ADH1B蛋白表达水平显著低于癌旁正常组织(P<0.001);预后良好组患者的ADH1B蛋白表达水平显著高于预后不良组(P<0.05);ADH1B低表达可缩短患者的无复发生存期(relapse-free survival,RFS)和总生存期(overall survival,OS)(均P<0.01);ADH1B表达水平与AFP、Ki67水平呈负相关趋势(均P<0.01)。Cox回归模型结果显示,ADH1B低表达是影响VI-HCC患者术后RFS和OS的独立危险因素。细胞学实验结果发现过表达ADH1B可抑制细胞的增殖、侵袭、迁移能力,促进其凋亡能力(均P<0.05)。结论肝脏代谢-解毒相关基因ADH1B在HCC中表达下调且其低表达缩短VI-HCC患者的RFS和OS,可能的机制是ADH1B抑制HCC细胞的增殖、侵袭和迁移的能力,促进HCC细胞的凋亡能力。Objective To investigate the clinical value of alcohol dehydrogenase 1B(ADH1B)in patients with hepatocellular carcinoma with vascular invasion(VI-HCC)and its effect on the biological function of liver cancer cells.Methods RNA sequencing of VI-HCC tissues,and GO,KEGG,GSEA,WGCNA analysis were performed.The expression level of ADH1B protein in tissues was detected by immunohistochemistry.The Kaplan-Meier method was used to draw the survival curve and the log-rank test was performed to analyze the survival difference of patients with different ADH1B protein expression groups.Univariate and multivariate Cox regression models were used to analyze the independent risk factors affecting the prognosis of patients with VI-HCC after surgery,and Spearman test was used to analyze the correlation between ADH1B and AFP/Ki67.The lentivirus overexpressing ADH1B and its negative control were used to infect the Huh7 cells to construct a stable ADH1B overexpression strain.The expression level of ADH1B mRNA in cells were detected by qRT-PCR.EDU and plate cloning assay were used to detect the proliferation ability of the cells.Transwell invasion and migration assay and scratch assay were used to detect the invasion and migration ability of the cells.Flow cytometry was used to detect apoptosis of cells.Results The results of RNA sequencing showed that the primary tumors of VI-HCC patients with poor prognosis after radical resection was characterized by metabolic reprogramming.The results of immunohistochemistry showed that the expression level of ADH1B protein in VI-HCC tissues was significantly lower than that in adjacent normal tissues(P<0.001).The expression of ADH1B protein in patients with good prognosis was significantly higher than that in patients with poor prognosis(P<0.05).Low expression of ADH1B could reduce both the relapse-free survival(RFS)and the overall survival(OS)of patients(all P<0.01).The expression level of ADH1B was negatively correlated with AFP and Ki67 levels(all P<0.01).Cox regression model showed that low exp

关 键 词：肝细胞癌 ADH1B 脉管侵犯 增殖 侵袭 迁移 预后 

分 类 号：R735.7[医药卫生—肿瘤]