肾细胞癌中候选生物标志物TCF21甲基化的研究及其意义

发布时间：2018-03-16 01:25

本文选题：DNA甲基化　切入点：肾细胞癌　出处：《福建医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：通过检测肾细胞癌组织和尿液中TCF21基因甲基化情况，分析TCF21基因甲基化水平与肾细胞癌临床特征的关系及相关性，用ROC曲线比较组织切片与尿液样本两种检测手段诊断肾细胞癌的灵敏度和特异度，探讨TCF21作为肾细胞癌中候选生物标志物在肾细胞癌早期诊断与预后的意义。方法：选取肾细胞癌组织55例作为试验组、随机选取相应远离肿瘤的正常肾组织21例作为对照组，同时随机选取肾细胞癌患者术前尿液标本33例及正常人尿液15例作为对照组。使用焦磷酸测序技术对上述样本的TCF21基因启动子甲基化水平进行定量检测。运用Mann-Whitney U非参数秩和检验统计分析，，分析TCF21甲基化水平与临床病理特征之间的关系；用Spearman分析TCF21甲基化水平与临床病理特征之间的相关性；用受试者接收操作特征(ROC)曲线来测绘真阳性率和假阳性率，分析比较肾细胞癌组织与尿液样本TCF21甲基化水平用于肾细胞癌诊断效价。结果： 1.肾细胞癌组织中TCF21基因甲基化水平明显高于癌旁正常肾组织中TCF21基因甲基化水平,二者的差异有统计学意义(P0.05)。肾细胞癌组织中TCF21甲基化水平与不同的年龄、吸烟史、病理分级及临床分期有关（P＜0.05），与年龄（R=0.166，P=0.002）、吸烟史（R=0.321，P=0.017）和Fuhrman分级（R=0.271，P=0.045）呈正相关。 2.肾细胞癌尿液患者中TCF21基因甲基化水平明显高于正常尿液样本中TCF21基因甲基化水平,二者的差异有统计学意义(P0.05)；尿液样本中TCF21甲基化水平与不同的肿瘤大小、病理分级及临床分期有关（P＜0.05），与肿瘤大小（R=0.622，P=0.000）、Fuhrman分级（R=0.441，P=0.010）和临床分期（R=0.411，P=0.017）呈正相关。 3.肾细胞癌组织中TCF21甲基化水平诊断肾细胞癌的ROC曲线下面积高于尿液样本中TCF21甲基化水平，肾细胞癌组织中TCF21甲基化水平和尿液样本中TCF21甲基化水平检测在诊断肾细胞癌时的ROC曲线下面积间的差异具有统计学意义（P＜0.05），组织切片诊断灵敏度和特异度为89%和61.9%，尿液样本诊断灵敏度和特异度分别为79%和100%。结论:TCF21与肾细胞癌的发生有关,肾细胞癌中的TCF21甲基化水平可以作为肾细胞癌早期诊断指标。TCF21甲基化水平与肾细胞癌的发展,恶性程度和分化程度有重要的关系。尿液样本检测具有收集方便、无创性及检测精确等优点，尿液样本的无创性高通量甲基化检测可作为组织甲基化检测的重要补充，可作为判断肾细胞癌恶性程度及预后等生物学的行为的重要检测手段。
[Abstract]:Objective: to study the relationship between methylation of TCF21 gene and clinical features of renal cell carcinoma (RCC) by detecting the methylation of TCF21 gene in renal cell carcinoma (RCC) tissues and urine. ROC curves were used to compare the sensitivity and specificity of tissue sections and urine samples in the diagnosis of renal cell carcinoma. The significance of TCF21 as a candidate biomarker in the early diagnosis and prognosis of renal cell carcinoma was discussed. Methods: 55 cases of renal cell carcinoma (RCC) were selected as experimental group and 21 cases of normal renal tissue which were away from tumor were randomly selected as control group. At the same time, 33 urine samples from patients with renal cell carcinoma and 15 normal urine samples were randomly selected as control group. The methylation level of TCF21 gene promoter was quantitatively detected by pyrosequencing technique. Mann-Whitney was used to detect the methylation level of the promoter of the TCF21 gene. U nonparametric rank sum test statistical analysis, To analyze the relationship between TCF21 methylation level and clinicopathological features; to analyze the correlation between TCF21 methylation level and clinicopathological features by Spearman; to map true positive rate and false positive rate by using receiver operating characteristics curve. To analyze and compare the TCF21 methylation level in renal cell carcinoma (RCC) tissues and urine samples for the diagnosis of renal cell carcinoma (RCC). Results:. 1. The methylation level of TCF21 gene in renal cell carcinoma tissues was significantly higher than that in adjacent normal renal tissues, and the difference was statistically significant (P 0.05). The methylation level of TCF21 in renal cell carcinoma tissues was different from that in different ages and smoking history. There was a positive correlation between pathological grade and clinical stage (P < 0.05) and age (P < 0.05), and was positively correlated with age, smoking history and smoking history (P < 0.017) and Fuhrman grade (P < 0.045). 2. The methylation level of TCF21 gene in urine samples of renal cell carcinoma was significantly higher than that in normal urine samples, and the difference between them was statistically significant (P 0.05), and the methylation level of TCF21 in urine samples was different from that of different tumor sizes. There was a positive correlation between pathological grade and clinical stage (P < 0.05), and was positively correlated with the size of the tumor (P < 0.010) and the clinical stage (P < 0.017), and was positively correlated with the size of the tumor (P < 0.05) and the Fuhrman grade (P < 0.010) and the clinical stage (P < 0.05). 3.The area under the ROC curve of TCF21 methylation in renal cell carcinoma was higher than that in urine samples. The difference of TCF21 methylation level in renal cell carcinoma tissue and TCF21 methylation level in urine samples was statistically significant (P < 0.05). The diagnostic sensitivity and specificity of tissue sections were 89% and 89% respectively. The diagnostic sensitivity and specificity of urine samples were 79% and 100, respectively. Conclusion the level of TCF21 methylation in renal cell carcinoma may be used as an early diagnostic marker of renal cell carcinoma, and the level of TCF21 methylation may be associated with the development of renal cell carcinoma. There is an important relationship between malignancy and differentiation. Urine sample detection has the advantages of convenient collection, noninvasive and accurate detection. The noninvasive high-throughput methylation detection of urine samples can be used as an important supplement to tissue methylation detection. It can be used as an important method to determine the malignant degree and prognosis of renal cell carcinoma (RCC).
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.11

【参考文献】