肝癌术后肝外转移相关DNA拷贝数变异分子标志初探

发布时间：2018-03-01 07:06

本文关键词： 肝细胞癌肝外转移微阵列比较基因组杂交拷贝数变异　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的肝细胞癌(hepatocellular carcinoma,HCC)是国内常见的恶性肿瘤,手术仍是目前HCC的主要根治性治疗方法,而术后肝外转移是患者预后差的一个重要因素。但目前对于具有何种分子特征的HCC易发生肝外转移仍知之甚少,因此筛选HCC术后肝外转移相关分子标志具有重要意义。既往研究显示DNA拷贝数变异(copy number aberrations,CNAs)与多种肿瘤进展和预后有关,且与HCC术后肝内复发相关,但CNAs与HCC术后远处转移的关系仍不清楚。本研究旨在探讨HCC术后肝外转移相关的CNAs分子标志。方法以66例HCC术后患者为研究对象,随访首次出现肝外转移的时间,无肝外转移生存时间定义为手术至出现肝外转移或末次随访之时间间隔。采用高分辨率Agilent 244K微阵列比较基因组杂交(array-based comparative genomic hybridization,a CGH)技术检测HCC组织基因组DNA的CNAs。采用Agilent G2565BA DNA微阵列扫描仪对杂交芯片进行扫描,运用Feature Extraction v9.5进行数据提取与分析。全基因组CNAs与HCC术后远处转移的相关性分析采用Cox风险比例模型和Log-rank检验,以Kaplan-Meier绘制生存曲线。P0.05为差异有统计学意义。统计学分析采用Stata 13.0统计学软件包(Stata Corporation,College Station,TX)。结果1.全组66例HCC样本的术后随访时间为1.6~90.5个月,中位随访时间39.9个月,总随访时间2633.4人月,随访期间共有25例(37.9%)患者出现肝外转移。2.单因素Cox分析显示,5个临床病理学因素与HCC术后肝外转移显著相关(均P0.05),其中高血压史、无完整肿瘤包膜、高TNM分期(Ⅲ期)为危险因素,而低血小板计数(100×109/L)和凝血酶原时间延长(12秒)为保护因素。3.全基因组CNAs与HCC术后远处转移的单因素Cox分析显示,5个CNAs片段是肝外转移的显著相关因素(均P0.05),其中6p21.32增益、15q11.2增益和20q12-13.13增益是HCC术后肝外转移的危险因素;4q12丢失和4q28.1-35.2丢失是转移保护因素。4.HCC术后肝外转移相关因素的多元逐步Cox回归分析结果显示,6p21.32增益、4q28.1-4q35.2缺失、高TNM分期(Ⅲ期)和高血压史是HCC术后远处转移的独立预后因素(均P0.05),其中6p21.32增益、高TNM分期和高血压史是转移危险因素,而4q28.1-4q35.2缺失是转移保护因素。5.CNAs组合与HCC术后肝外转移的相关性:依据6p21.32增益与4q28.1-35.2丢失状态优化组合,发现6p21.32增益、6p21.32无增益/4q28.1-35.2丢失和6p21.32无增益/4q28.1-35.2无丢失3组HCC的术后无肝外转移生存差异有统计学意义(Log-rank检验,P0.01);相比6p21.32无增益/4q28.1-35.2无丢失组,6p21.32无增益/4q28.1-35.2丢失组的肝外转移风险显著降低至0.18倍(95%CI=0.06~0.57,P0.01),而6p21.32增益组的肝外转移风险显著增加至2.91倍(95%CI=1.13~7.46,P0.05)。结论1.6p21.32增益、15q11.2增益和20q12-13.13增益是HCC术后肝外转移的危险因素,而4q12丢失和4q28.1-35.2丢失是转移保护因素。2.6p21.32增益和4q28.1-35.2丢失是HCC术后肝外转移的独立预后因素,前者是转移危险因素,后者是转移保护因素。3.基于6p21.32增益和4q28.1-35.2丢失2个独立预后因素的CNAs优化组合可在手术早期对HCC术后肝外转移风险作出预判,从而为临床治疗决策提供依据。
[Abstract]:Objective hepatocellular carcinoma (hepatocellular, carcinoma, HCC) is a common malignant tumor in China, surgery is still the main HCC radical treatment methods, and postoperative liver metastasis is an important factor of poor prognosis. But for what is the molecular characteristics of HCC susceptible to extrahepatic metastasis remains poorly understood. The screening of HCC postoperative extrahepatic metastasis related molecular markers is important. Previous studies have shown that the copy number variation of DNA (copy number aberrations, CNAs) associated with the progression and prognosis of various tumors, and is associated with intrahepatic recurrence after HCC, but the relationship between CNAs and metastasis after HCC remains unclear. The purpose of this study is to to investigate the HCC postoperative extrahepatic metastasis CNAs molecular marker related. Methods 66 cases of HCC patients were followed up for the first time as the research object, extrahepatic metastasis, extrahepatic metastasis survival time is defined as to the liver surgery. Shift or last follow-up time interval. By using high resolution Agilent 244K microarray comparative genomic hybridization (array-based comparative genomic hybridization, a CGH DNA HCC) in detecting genomic organization technology using CNAs. Agilent G2565BA DNA microarray scanner on hybrid chip scan, using Feature Extraction v9.5 data extraction and analysis of genomic CNAs and correlation analysis. After HCC of distant metastasis with Cox proportional hazard model and Log-rank test, Kaplan-Meier.P0.05 to draw survival curves for significant difference. Statistical analysis using Stata 13 statistical software package (Stata Corporation, College Station, TX). All 66 cases were followed up for 1. HCC sample results after operation for 1.6~90.5 months. After a median follow-up of 39.9 months, the total follow-up time of 2633.4 months follow-up period, a total of 25 cases (37.9%) patients with liver. Shift.2. Cox univariate analysis showed that 5 factors of clinical pathology and HCC postoperative extrahepatic metastasis was significantly correlated (P0.05), the history of hypertension, no intact tumor capsule, high TNM stage (stage III) as a risk factor, and low platelet count (100 x 109/L) and prothrombin time (12 a second) appear as single factors of distant metastasis protective factors of.3. genomic CNAs and HCC after Cox analysis, 5 CNAs fragments were significantly related with extrahepatic metastasis (P0.05), the 6p21.32 gain, 15q11.2 gain and 20q12-13.13 gain are risk factors of HCC postoperative extrahepatic metastasis; 4q12 loss and 4q28.1-35.2 loss multiple factors related to transfer of protective factors of.4.HCC postoperative extrahepatic metastasis by Cox regression analysis showed that the 6p21.32 gain, 4q28.1-4q35.2 loss, high TNM stage (stage III) and history of hypertension is an independent prognostic factor for distant metastasis after HCC (P0.05), of which 6 P21.32 high gain, TNM staging and history of hypertension were the risk factors of metastasis, metastasis and 4q28.1-4q35.2 deletion is a protective factor for.5.CNAs in combination with HCC postoperative extrahepatic metastasis: Based on the 6p21.32 gain and 4q28.1-35.2 lost state optimization, 6p21.32 gain, 6p21.32 no gain without liver loss of /4q28.1-35.2 and 6p21.32 gain /4q28.1-35.2 without loss of 3 HCC the postoperative metastasis survival difference was statistically significant (Log-rank, P0.01); no gain no loss compared to 6p21.32 /4q28.1-35.2 group, 6p21.32 /4q28.1-35.2 group of liver abversion gain loss risk shift significantly reduced to 0.18 times (95%CI=0.06~0.57, P0.01), and the risk of 6p21.32 gain group extrahepatic metastasis significantly increased to 2.91 times (95%CI=1.13~7.46, P0.05). Conclusion 1.6p21.32 gain, 15q11.2 gain and 20q12-13.13 gain are risk factors of HCC postoperative extrahepatic metastasis, and loss of 4q12 and 4q28.1-35. 2 transfer loss is a protective factor for.2.6p21.32 gain and loss of 4q28.1-35.2 were independent prognostic factors of HCC postoperative extrahepatic metastasis, the former is the transfer of risk factors, the latter is the transfer of protective factors.3. optimization can be in operation early on HCC postoperative extrahepatic metastasis risk to predict CNAs 6p21.32 gain and 4q28.1-35.2 lost 2 independent prognostic factors based on, so as to provide the basis for clinical decision making.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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