微卫星位点杂合性缺失检测鉴定多结节肝细胞癌克隆起源的方法及其临床意义

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目的:建立微卫星位点杂合性缺失(LOH)检测多结节肝细胞癌克隆起源的技术并评估临床意义。方法:选择2015年1月至2018年12月于福建医科大学孟超肝胆医院肝胆外科进行手术的25例多结节肝细胞癌患者作为研究对象,其中男性15例,年龄(52.74±7.60)岁,收集患者的临床病理学特征,检测肿瘤结节微卫星位点缺失情况。利用多重荧光聚合酶链反应(PCR)技术对患者DNA的10个微卫星位点进行扩增并进行片段分析。随后,根据微卫星位点杂合性缺失检测的判断标准,将多结节肝细胞癌患者分为肝内转移组(IM)以及多中心生发组(MO)。最后,分析两组患者肿瘤在组织学特征、一般临床资料、血清学指标以及生存时间上存在的差异。应用SPSS 20.0统计软件分析,对一般临床资料的差异采用两独立样本n t检验或者n χ2检验。n 结果:8例患者被判断为肝内转移。单因素分析结果表明,IM组存在脉管癌栓的明显高于MO组(41.18%比25.00%,n χ2=4.590,n P<0.05),差异有统计学意义;IM组的肝炎的Gn 2~Gn 4分期的比例小于MO组(0.00%比52.94%,n χ2=6.618,n P<0.05),差异有统计学意义;IM的肝硬化等级低于MO组(37.50%比88.89%,n χ2=7.434,n P<0.05),差异有统计学意义。同时,Kaplan-Meier生存曲线则提示MO组患者的预后好于IM组(37个月比14个月,n χ2=7.232,n P<0.05),差异有统计学意义。n 结论:LOH检测技术可鉴别多结节肝细胞癌的克隆起源,且可指导肝细胞癌患者的临床治疗及预后评估。“,”Objective:To establish techniques for heterozygous loss of microsatellite on detecting the origins of multiple hepatocellular carcinomas.Methods:Totally, 25 patients with multi-nodular hepatocellular carcinoma who underwent surgery in Mengchao Hepatobiliary Hospital of Fujian Medical University from January 2015 to December 2018 were enrolled. Among them they were 15 males, with an average age of (52.74±7.60) years old.The clinical pathological characteristics of the study subjects were collected and the absence of microsatellite sites in tumor nodules was also measured. The DNA of the cancer tissues and adjacent tissues was extracted. The 10 microsatellite loci were amplified using multiple fluorescent polymerase chain reaction (PCR) technology, and then analyzed by fragment analysis method. According to the criteria of the loss of heterozygosity of microsatellite, the patients with multinodular hepatocellular carcinoma were divided into the intrahepatic metastasis (IM) and multicentric occurrence (MO) groups. Finally, the differences in histological characteristics, general clinical data, serological indicators and survival time between the two groups were also compared.Results:Eight of 25 patients were judged to be IM. Univariate analysis showed that the proportion of vascular tumor thrombi in the IM group was significantly higher than that in the MO group (41.18% vs. 25.00%, n χ2=4.590, n P<0.05). The proportion of grading 2-grading 4 (Gn 2-Gn 4) stages of hepatitis in the IM group was lower than that in the MO group (0.00% vs. 88.89%, n χ2=6.618, n P<0.05). The cirrhosis grade in the IM group was lower than that in the MO group (37.50% vs. 88.89%,n χ2=7.434, n P<0.05). In addition, the Kaplan-Meier survival curve further indicated that the prognosis of the MO group was better than that of the IM group (34 months vs. 14 months,n χ2=7.232, n P<0.05).n Conclusion:Detecting the loss of heterozygosity of microsatellite could be used to identify clonal origins of multiple hepatocellular carcinomas, which could guide the clinical treatment and prognosis evaluation of patients with hepatocellular carcinoma.
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