肝细胞癌病人血浆循环DNA定量分析及其临床意义

Quantitative Analysis of Circulating DNA Level in Plasma from Patients with Hepatocellular Carcinoma and Its Potential Clinical Value

目的 定量检测肝细胞癌 (HCC)病人血浆循环DNA并探讨其临床价值。方法 收集 79例肝功能正常的HCC病人术前血浆样本以及 2 0例健康志愿者、2 0例代偿期肝硬化病人和 2 0例活动性乙型肝炎病人的血浆样本 ,抽提血浆循环DNA。将血浆循环DNA与荧光染料SYBRgreenⅠ按比例稀释并充分混匀后通过紫外与可见光成像分析系统定量。结果 HCC、肝硬化和肝炎病人血浆循环DNA浓度分别为 (47.1± 4 3.7)ng/mL、(30 .0± 1 3.3)ng/mL和 (6 5 .3± 71 .6 )ng/mL ,均明显高于健康人群血浆循环DNA浓度 (1 7.6± 9.5 )ng/mL ,差异在统计学上有显著性 (P =0 .0 0 0 ;P =0 .0 0 2 ;P =0 .0 0 1 ) ,而HCC和肝硬化病人以及病毒性肝炎病人两组之间的血浆循环DNA浓度无明显差异 (P =0 .1 91 ;P =0 .6 85 )。伴肝内播散灶或脉管癌栓的 4 1例HCC病人的血浆循环DNA浓度 (5 4 .3± 4 1 .6 )ng/mL明显高于不伴肝内播散灶或脉管癌栓的 38例病人 (39.3± 4 5 .0 )ng/mL(P=0 .0 0 2 )。血浆循环DNA浓度与HCC病人肿瘤大小、TNM分期、2年无瘤生存率、1年和 2年总体生存率密切相关 (P =0 .0 0 8;P =0 .0 4 0 ;P =0 .0 1 0 ;P =0 .0 1 6 ;P =0 .0 0 1 ) ,而与肿瘤数目、肿瘤包膜、肿瘤分化、血清AFP浓度及 1年无瘤生存率无明显关系。多因素分?

Purpose To quantify the circulating tumor DNA in plasma from patients with hepatocellular carcinoma (HCC) and evaluate its potential clinical value. Methods Blood samples were collected from 79 HCC patients with normal liver function preoperatively,and from 20 patients with compensated liver cirrhosis,20 patients with hepatitis B and 20 healthy volunteers.Plasma DNA was extracted and analyzed by ultraviolet transilluminator system.The circulating plasma DNA level was calculated by regression equation which was made by intensity of standard DNA content. Results Significant differences (P=0.000;P= 0.002;P=0.001) in mean circulating plasma DNA levels were found between healthy volunteers (17.6±9.5)ng/mL and patients with HCC (47.1±43.7)ng/mL,with liver cirrhosis (30.0±13.3) ng/mL or with hepatitis (65.3±71.6) ng/mL.However,there was no significant difference in the mean circulating plasma DNA levels between patients with HCC and liver cirrhosis (P=0.191) or between patients with HCC and hepatitis (P=0.586).Among the HCC patients,the mean circulating plasma DNA level of patients with intrahepatic spreading or vascular invasion (54.3±41.6)ng/mL was significantly higher than that of those without intrahepatic spreading and vascular invasion (39.3±45.0)ng/mL(P=0.002).Circulating plasma DNA level was found to be closely associated with tumor size,TNM stage,2-year disease-free survival (DFS),1-year and 2-year overall survival (OS) (P=0.008; P=0.040;P= 0.010;P=0.016;P=0.001),but not associated with tumor number,tumor capsule,tumor differentiation,serum AFP level or 1-year DFS.Multivariate analysis showed high plasma DNA level was independently associated with decreased OS (P=0.020). Conclusions Circulating plasma DNA level has a limited va-lue in the diagnosis of HCC.However,it can initially predict the metastasis potential and prognosis of those diagnosed HCC patients before operation,which may help to choose therapeutic approach.