Objective: To explore the diagnostic value of the serum metabolites identified by high-performance liquid chromatography-mass spectrometry (HPLC/MS) for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).
Methods: A total of 126 patients admitted to Tianjin Third Central Hospital were enrolled, including 27 patients with HBV-related hepatitis with negative viral DNA (DNA-N), 24 with HBV-related hepatitis with positive viral DNA, 24 with HBV-related liver cirrhosis, 27 with HBV-related HCC undergoing surgeries or radiofrequency ablation, and 24 with HBV-related HCC receiving interventional therapy, with 25 healthy volunteers as the normal control group. Serum samples were collected from all the subjects for HPLC/MS analysis, and the data were pretreated to establish an orthogonal partial least- squares discriminant analysis (OPLS-DA) model. The differential serum metabolites were preliminarily screened by comparisons between the HBV groups and the control group, and the characteristic metabolites were identified according to the results of non-parametric test. The potential clinical values of these characteristic metabolites were evaluated using receiver operator characteristic curve (ROC) analysis.
Results: A total of 25 characteristic metabolites were identified in the HBV- infected patients, including 9 lysophosphatidylcholines, 2 fatty acids, 17α-estradiol, sphinganine, 5-methylcytidine, vitamin K2, lysophosphatidic acid, glycocholic acid and 8 metabolites with few reports. The patients with HBV- related HCC showed 22 differential serum metabolites compared with the control group, 4 differential metabolites compared with patients with HBV-related liver cirrhosis; 10 differential metabolites were identified in patients with HBV-related HCC receiving interventional therapy compared with those receiving surgical resection or radiofrequency ablation. From the normal control group to HBV-related HCC treated by interventional therapy, many metabolites underwent variations following a similar pattern.
Conclusions: We identified 25 characteristic metabolites in patients with HBV-related HCC, and these metabolites may have potential clinical values in the diagnosis of HBV-related HCC. The continuous change of some of these metabolites may indicate the possibility of tumorigenesis, and some may also have indications for the choice of surgical approach.
目的: 通过HPLC/MS平台探索血清代谢物对于乙肝相关性肝细胞癌（HCC）的诊断价值。
方法: 选取126例就诊于天津市第三中心医院住院患者，包括27例乙肝DNA阴性乙肝肝炎患者、24例乙肝DNA阳性乙肝肝炎患者、24例乙肝肝硬化患者、27例乙肝相关性HCC行手术/射频治疗患者以及24例乙肝相关性HCC行介入治疗患者，另选取查体中心健康志愿者25例为对照组。将血清样本上机所得数据进行前处理，构建正交偏最小二乘判别分析（OPLS-DA）模型，基于乙肝组和正常对照组进行代谢物初筛。根据非参数检验的结果，最终筛选和鉴定特征代谢物。利用ROC分析这些特征代谢物的临床潜在应用价值。
结果: 筛选并鉴定出25种特征代谢物，包括9种溶血性磷脂酰胆碱、2种脂肪酸、17 α-雌二醇、二氢神经鞘氨醇、5-甲基胞苷、维生素K2、溶血磷脂酸、甘氨胆酸以及报道较少的8种物质。与正常对照组相比，22种代谢物在乙肝相关性HCC组中存在差异表达。与乙肝肝硬化组相比，4种代谢物在乙肝相关性HCC组中存在差异表达。与乙肝相关性HCC行手术/射频治疗组相比，10种代谢物在乙肝相关性HCC行介入治疗组中存在差异表达。从正常对照组到乙肝相关性HCC行介入治疗组，许多代谢物呈相同的变化趋势。
结论: 基于LC/MS平台，本研究成功地构建疾病区分模型并筛选和鉴定出25种特征代谢物，这些代谢物对乙肝相关性HCC具有一定诊断和分期效能。有些特征代谢物呈连续性的变化预示着癌变的可能；有些代谢物对手术方式的选择也具有指导意义。
Keywords: hepatitis; hepatitis B virus; hepatocellular carcinoma; liver cirrhosis; metabonomics.