【作者】 黄芸；

【导师】 杨文龙；

【作者基本信息】 南昌大学 ， 内科学（传染病学）（专业学位）， 2022， 硕士

【摘要】 目的:探讨尿液RAS相关区域家族1A(RASSF1A)基因甲基化水平与甲胎蛋白(AFP)单独及联合检测在乙肝(HBV)相关性肝细胞癌中的诊断效能及临床价值。方法:收集2019年7月至2021年1月在南昌大学第二附属医院就诊患者145例,分为4组,其中HBV相关性肝细胞癌患者58例设为肝癌组,同期门诊或住院的肝硬化患者28例设为肝硬化组,肝炎患者29例设为肝炎组,健康体检者30例设为健康组。分别测定各组血清AFP、尿液RASSF1A甲基化水平并收集患者临床基线资料,分析AFP、RASSF1A甲基化水平及在上述各组间的表达水平差异。同时比较二者在不同肿瘤负荷状态下(肿瘤的单发与多发、癌瘤直径大小、淋巴结转移以及是否合并肝硬化)的表达差异;比较RASSF1A甲基化在不同AFP浓度分组中的表达水平差异。采用多因素回归分析法确立独立变量,并利用二元logistics回归分析法构建诊断模型,绘制受试者工作(ROC)曲线分析AFP、RASSF1A甲基化单独及联合检测在肝细胞癌(HCC)的诊断效能。结果:1.尿液RASSF1A甲基化及AFP在肝癌组中表达水平均显著高于肝硬化、肝炎及健康组(P<0.05);2.尿液RASSF1A甲基化水平在不同AFP浓度HCC患者中表达无明显差异(P>0.05);3.在不同肿瘤负荷状态下,AFP在多发肝癌组表达水平显著高于单发肝癌组(P<0.05),在有淋巴结转移组中显著高于无淋巴结转移组(P<0.05),在合并肝硬化组及单纯肝癌组中表达无显著差异(P>0.05),与肿瘤直径大小之间具有弱相关性(P<0.05);而尿液RASSF1A甲基化水平在上述各分组间均无明显差异(P>0.05),且与肝癌肿瘤直径大小间无显著相关性(P>0.05)。4.尿液RASSF1A甲基化水平诊断HBV相关性肝细胞癌的ROC曲线下面积AUC为0.743(95%CI 0.654-0.832),所对应的最佳截断值为3.735cp/m L,敏感性为70.7%,特异性66.7%;AFP的AUC为0.758(95%CI 0.669-0.848),曲线所对应的最佳截断值为171.85ng/m L,敏感性为56.9%,特异性为91.2%。AFP、m RASSF1A联合诊断模型的AUC为0.848(95%CI 0.780-0.915),敏感性为86.2%,特异性为81.3%。两指标结合年龄和性别模型的AUC为0.915(95%CI 0.866-0.964),敏感性为82.8%,特异性为98.6%。5.尿液RASSF1A甲基化在AFP阴性HBV相关性HCC中的ROC曲线下面积AUC为0.707(95%CI 0.582-0.832);在AFP表达低水平的HBV相关性HCC中的ROC曲线下面积AUC为0.719(95%CI 0.582-0.832)。6.尿液RASSF1A甲基化在早期HBV相关性肝细胞癌诊断中的ROC曲线下面积AUC为0.715(95%CI 0.593-0.837);AFP在早期HBV相关性肝细胞癌诊断中的ROC曲线下面积AUC为0.574(95%CI 0.424-0.723),两指标联合诊断模型的AUC为0.754(95%CI 0.639-0.870)。结论:1.尿液RASSF1A基因甲基化在HBV相关性HCC诊断中具有一定潜能;与AFP联合检测的诊断效能优于单独AFP检测。并且在低水平AFP、AFP阴性及早期HBV相关性HCC患者中仍具有良好的诊断效能。2.尿液RASSF1A基因甲基化表达与AFP水平、肝癌的多发与单发、肿瘤直径大小及淋巴结转移、是否合并肝硬化等临床特征无关。更多还原

【Abstract】 Objectives:To investigate the diagnostic efficacy and clinical value of urinary RAS-related region family 1A(RASSF1A)gene methylation levels and alpha-fetoprotein(AFP)detection alone or in combination in hepatitis B(HBV)-related hepatocellular carcinoma.Methods:A total of 145 patients who visited the Second Affiliated Hospital of Nanchang University from July 2019 to January 2021 were selected and divided into4 groups.Among them,58 patients with HBV-related hepatocellular carcinoma were selected as the liver cancer group,and 28 patients with liver cirrhosis who were outpatient or hospitalized during the same period were selected as the liver cirrhosis group,29 hepatitis patients were set as hepatitis group,and 30 healthy subjects were set as healthy group.The methylation levels of serum AFP and urine RASSF1 A in each group were measured respectively,and the clinical baseline data of the patients were collected.At the same time,the expression differences of the two groups were compared under the status of tumor burden(single and multiple tumors,tumor diameter,lymph node metastasis,and liver cirrhosis);the expression levels of RASSF1 A methylation at different AFP concentrations were compared.Multivariate regression analysis was used to establish independent variables,and binary logistic regression analysis was used to construct a joint diagnostic model,and receiver operating(ROC)curves were drawn to analyze the detection of AFP and RASSF1 A methylation alone and in combination in hepatocellular carcinoma(HCC).diagnostic performance.Results:1.The levels of RASSF1 A methylation and AFP in the urine of the hepatocellular carcinoma group were significantly higher than those of the liver cirrhosis,hepatitis and healthy groups,and the difference was statistically significant(P<0.05);2.There was no significant difference in the expression of urinary RASSF1 A methylation among HCC patients in different AFP concentration groups(P>0.05);3.Under different tumor burden status,the expression level of AFP in the multiple hepatocellular carcinoma group was significantly higher than that in the single hepatocellular carcinoma group(P<0.05),and in the group with lymph node metastasis was significantly higher than that in the group without regional lymph node metastasis(P<0.05).The combined liver cirrhosis group was significantly higher than the simple liver cancer group(P < 0.05),and there was a weak correlation with tumor diameter(P < 0.05);the level of urinary RASSF1 A methylation had no significant difference among the above groups(P>0.05).There was no significant correlation between urinary RASSF1 A methylation and hepatocellular carcinoma tumor diameter(P>0.05).4.The area under the ROC curve of urine RASSF1 A methylation level in the diagnosis of HBV-related hepatocellular carcinoma was 0.743(95%CI 0.654-0.832),the corresponding optimal cutoff value was 3.735,the sensitivity was 70.7%,and the specificity was 70.7%.The AUC of AFP was 0.758(95%CI 0.669-0.848),the best cut-off value corresponding to the curve was 171.85ng/m L,the sensitivity was 56.9%,and the specificity was 91.2%.The AUC for the combined diagnosis of AFP and m RASSF1 A was 0.848(95%CI 0.780-0.915),with a sensitivity of 86.2% and a specificity of 81.3%.The AUC of the two-indicator combined age and gender model was 0.915(95% CI 0.866-0.964),with a sensitivity of 82.8% and a specificity of98.6%.5.The area under the ROC curve of urinary RASSF1 A methylation in AFP-negative liver cancer was 0.707(95%CI 0.582-0.832);in low-level AFP liver cancer,the area under the ROC curve AUC was 0.719(95%CI 0.582-0.832).6.The area under the ROC curve of urine RASSF1 A methylation in the diagnosis of early HBV-related hepatocellular carcinoma was 0.715(95%CI0.593-0.837);the ROC curve of AFP in the diagnosis of early HBV-related hepatocellular carcinoma The area AUC was 0.574(95%CI 0.424-0.723),and the AUC of the two-indicator combined diagnostic model was 0.754(95%CI0.639-0.870).Conclusions:1.Urine RASSF1 A gene methylation has a certain potential in the diagnosis of HBV-related HCC;the diagnostic performance of combined detection with AFP is better than that of single AFP detection.And it still has good diagnostic performance in low-level AFP,AFP-negative and early-stage HBV-related HCC patients.2.The methylation expression of RASSF1 A gene in urine was not related to the clinical characteristics of AFP level,multiple or single hepatocellular carcinoma,tumor diameter,lymph node metastasis,and liver cirrhosis.更多还原