应用克隆分析鉴别肝癌多中心发生与肝内转移

摘要

目的 探讨克隆分析对肝癌多中心发生(multicentric occurrence,MO)与肝内转移(intrahepatic metastasis,IM)的鉴别意义.方法 用微卫星多态性技术检测多癌灶肝细胞癌的杂合性缺失(loss of heterozygosity,LOH)和微卫星不稳定性(microsatellite instability,MSI),用Southern Blot法检测HBV整合位点,分析癌灶间的克隆来源,判断MO与IM,并与临床病理和影像学分析结果进行比较.结果 对35例多发肝癌病人79个瘤结节与非癌组织进行LOH和MSI检测,5例(14.3%)判断为MO,29例(82.9%)为IM,1例(2.9%)既存在MO又存在IM瘤灶.来自34例多发肝癌病人的77个瘤结节能够进行HBV整合位点分析,其中6例(17.6%)判断为MO,27例(79.4%)为IM,1例(2.9%)包含两种类型的瘤灶.两种方法所得分类结果具有显著的正相关关系(rs=0-909,P＜0.001),但是它们与临床病理及影像学结果无明显相关性(rs=0.133,P=0.468;rs=0.262,P=0.155).另外,在克隆分析确定的MO与IM病人中,MO组的复发时间明显晚于IM组(P=0.001).结论 使用微卫星多态性技术检测LOH和MSI,用Southem Blot法检测HBV整合位点,进而推测多癌灶肝癌间的克隆系来源,有助于鉴别MO与IM,从而指导临床治疗及预后评估.%Objective To explore the differential diagnostic significance of clone analysis for multicentric occurrence (MO) and intrahepatic metastasis (IM) in hepatocellular carcinomas (HCCs).Methods Loss of heterozygosity (LOH) and microsatellite instability (MSI) were analyzed by microsatellite polymorphism test and the integration sites of HBV were assessed by Southern blot in each nodule of the HCCs. The clonalities were then compared between each nodule of the same patient and the diagnosis of MO or IM was concluded. Finally, the results based on clonality analysis were compared with those according to clinicopathological and imaging data. Results According to the results of LOH and MSI in 79 nodules and nontumorous tissue from 35 cases of mutiple HCCs, 5 (14.3%)and 29 cases (82.9 %) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The integration sites of HBV could be analyzed in 77 nodules from 34 multiple HCCs. Among them, 6 (17. 6%) and 27 cases (79.4%) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The classification results of microsatellite polymorphism test and HBV integration sites analysis demonstrated a significant positive correlation (rs = 0.909, P＜0.001). Nevertheless, neither the classification of microsatellite polymorphism test nor that of HBV integrate sites analysis was correlated with the discrimination according to clinicopathologic and imaging data (rs=0. 133, P=0. 468, rs =0. 262, P=0. 155, respectively). Recurrence in patients in the MO group occurred significantly later than that in IM cases who were diagnosed by clonality analyses (P=0. 001). Conclusion The clonality analysis based on the results of LOH and MSI or assessments of HBV integrate sites is useful for the differential diagnosis of MO and IM and their treatment and prognosis.