EZH2、Bmi-1、P16、IMP-1和Survivin自身抗体在多种肿瘤中表达变化的研究

发布时间：2018-03-01 14:24

  本文关键词： 自身抗体 诊断标志物 肺癌 食管癌 乳腺癌 肝癌 肿瘤相关抗原　出处：《吉林大学》2016年博士论文　论文类型：学位论文

【摘要】：目的:中国是世界人口大国也是癌症高发的国家,由于肿瘤多起病隐匿、早期表现不明显,导致多数肿瘤患者在发现时就已经处于中晚期阶段,治疗效果不理想患者生活质量大大降低。所以肿瘤的早期诊断、早治疗对肿瘤患者来说十分重要。然而目前肿瘤的诊断主要依赖于影像学和细胞学检查,诊断能力有限,目前亟待出现一种能更早期诊断肿瘤的手段。自从Landsteier提出机体内存在自身抗体(autoantibody)以来,人们逐渐对自身抗体有了深刻的认识。在长期肿瘤发病机制研究过程中,学者们认为基因突变是最基本的触发环节或始动因素,这些异常表达的蛋白就成为了肿瘤抗原。相对于浓度较低的肿瘤抗原,自身抗体的检测具有更高的敏感性。因此我们检测高发肿瘤中自身抗体的表达变化,以期寻找到可用于肿瘤辅助诊断的新型肿瘤标志物。方法:通过查阅文献和课题组前期工作基础,本研究中纳入5种目标蛋白,分别是m RNA的结合蛋白IMP-1、凋亡抑制基因蛋白Survivin、肿瘤抑制因子P16、转录调节蛋白EZH2和原癌基因蛋白Bmi-1。利用免疫学分析软件和表位数据库,预测、设计并合成5种蛋白的多肽抗原片段,应用经过我们优化改良的ELISA检测方法,包被多肽抗原,根据免疫学抗原抗体结合原理检测血浆中自身抗体水平。研究共纳入未经任何抗癌治疗的首次诊断为恶性肿瘤的肝癌患者、食管癌患者、肺癌患者、乳腺癌患者和健康人群各200例。设置质控血衡量检测的稳定性,根据酶标仪测得的样本OD和质控OD计算相对结合指数,降低非特异结合和检测误差。应用SPSS21.0进行数据统计和分析,比较自身抗体水平在不同肿瘤、不同分期中的表达变化,分析其在不同肿瘤中表达特点的同时,探讨其在肿瘤诊断中的作用。结果:1肝癌患者血浆5种肿瘤相关抗原自身抗体检测中,EZH2和Bmi-1自身抗体水平出现显著升高(p0.001;p0.001),进一步按肿瘤分期分析发现,早期患者EZH2和Bmi-1自身抗体水平高于晚期患者。受试人群ROC曲线分析发现,EZH2和Bmi-1自身抗体检测在肝癌诊断中具有较高的应用价值,特异度90%情况下,肝癌检测敏感度分别为22%和32%。2在非小细胞肺癌患者血浆中,同样检测了EZH2、Bmi-1、P16、IMP-1和Survivin自身抗体表达变化。其中P16和Survivin自身抗体表达出现显著升高(p0.001;p0.001),虽然IMP-1亦出现升高趋势,但没有统计学差异。以肿瘤分期分层分析中,P16和Survivin自身抗体在肺癌早期升高更明显。受试人群ROC曲线分析发现,P16和Survivin自身抗体在肺癌诊断中具有很好的临床价值,特异度90%情况下其敏感度分别为32%和20%。3食管癌患者血浆中,我们同样检测了EZH2、Bmi-1、P16、IMP-1和Survivin自身抗体表达变化。食管癌和肺癌中自身抗体表达变化趋势相似,P16和Survivin自身抗体表达出现显著升高(p0.001;p0.001),以肿瘤分期分层分析中,P16和Survivin自身抗体在食管癌早期升高不如肺癌明显。受试人群ROC曲线分析发现,特异度90%情况下,食管癌检出的敏感度分别为26%和15%。4为更好判断肿瘤相关抗原自身抗体在不同肿瘤诊断中的特异性,在乳腺癌患者血浆中,我们同样检测了EZH2、Bmi-1、P16、IMP-1和Survivin自身抗体表达变化。P16和Survivin自身抗体表达在乳腺癌中略有升高,与健康组存在统计学差异(p=0.013;p=0.000),在肿瘤分期中没有明显的差异。受试人群ROC曲线分析发现,特异度90%情况下,乳腺癌检出的敏感度分别为20%和16%。5以肝癌、肺癌、食管癌和乳腺癌的临床传统标志物AFP、EGFR和HER2为分层表型,分析EZH2、Bmi-1、P16、IMP-1和Survivin自身抗体在不同表型表达变化发现,HER2和EGFR过表达与自身抗体表达无关。有趣的是,肝癌患者中AFP的表达与EZH2和Bmi-1自身抗体的表达呈现正相关。结论:1EZH2和Bmi-1自身抗体在肝癌患者中表达水平明显升高,而在其他肿瘤中表达变化并不明显,EZH2和Bmi-1自身抗体可能是肝癌的潜在生物学标志物,对肝癌诊断有一定辅助作用,且与AFP的表达有显著相关性。其联合检测可有效提高肝癌检出的敏感度和特异度。2非小细胞肺癌中P16和Survivin自身抗体表达水平在肺癌患者中明显升高,其中P16和Survivin自身抗体的敏感度和特异度最好,可以作为肺癌诊断指标,有待进一步优化。3食管癌中P16和Survivin自身抗体表达水平在肿瘤组和健康人群之间存在显著差异,但不如肺癌中更明显,可以作为食管癌辅助诊断的参考指标。4Survivin自身抗体表达在乳腺癌中略有升高,虽然Survivin自身抗体在肺癌和食管癌也均明显升高,但在以上3种肿瘤中其检测的敏感度存在差异,需进一步扩大样本来验证其作为标志物的特异性。5自身抗体检测在肿瘤早期诊断中具有一定辅助作用,有成为肿瘤诊断新方法的潜能。
[Abstract]:Objective: Chinese is the most populous country in the world is also a high incidence of cancer, because cancer onset and early performance is not obvious, resulting in the majority of cancer patients were found already in the late stage, the treatment effect is not ideal. So greatly reduced the quality of life of patients with cancer early diagnosis, early treatment is very important for cancer patients. However, the tumor diagnosis depends on the histological and cytological examination imaging, diagnosis ability is limited, at present urgently needed a more early diagnosis of tumors. Since Landsteier proposed the body autoantibodies (autoantibody) since the people began to have a deep understanding of autoantibodies in the pathogenesis of cancer. The long-term process of scholars that gene mutation is the most basic link or trigger initiating factors, the abnormal expression of the protein has become a tumor antigen for concentration. Tumor antigen is low, autoantibody detection has higher sensitivity. Therefore we detected expression in tumor autoantibodies, in order to search for new tumor markers for tumor diagnosis. Methods: through the literature and previous work foundation, into 5 kinds of target protein in this study respectively, is RNA m binding protein IMP-1, apoptosis gene Survivin, tumor suppressor P16, transcriptional regulatory protein EZH2 and oncogene protein Bmi-1. using immunological epitope analysis software and database, forecast, design and synthesis of peptide antigen fragment of 5 proteins, by using ELISA detection method we optimize the package is polypeptide antigen, antigen antibody binding principle according to the immunological detection of plasma levels of autoantibodies. The study included the first diagnosis for malignant tumor of liver cancer is not treated by any anticancer Patients, patients with lung cancer, esophageal cancer, breast cancer patients and healthy controls in all 200 cases. The detection of blood quality control measure of stability, according to the quality control samples OD and OD measured eliasa calculation of relative binding index, reduce the non-specific binding and error detection. The application of SPSS21.0 statistics and comparative analysis, autoantibodies the level of expression in different tumors in different stages, and to analyze its expression characteristics in different tumors, and explore its role in tumor diagnosis. Results: the detection of 1 liver cancer patients, 5 tumor associated antigens, EZH2 and Bmi-1 autoantibodies level increased significantly (p0.001; p0.001), further according to the tumor stage analysis found that early levels in patients with EZH2 and Bmi-1 antibodies was higher than that in patients with advanced human subjects. ROC analysis indicated that the detection of EZH2 and Bmi-1 antibodies with high in diagnosis of hepatocellular carcinoma The application value, the specificity was 90% cases, liver cancer detection sensitivity was 22% and 32%.2 in plasma of patients with non-small cell lung cancer were also detected, EZH2, Bmi-1, P16, IMP-1 and Survivin expression changes of autoantibodies. The expression of P16 and Survivin increased significantly in the presence of autoantibodies (p0.001; p0.001), although the IMP-1 also appeared increased, but there is no significant difference in tumor staging in stratified analysis, P16 and Survivin antibodies increased more significantly in lung cancer early. The subjects were ROC curve analysis, P16 and Survivin antibodies in the diagnosis of lung cancer has good clinical value, specificity 90% case sensitivity were 32% and 20%.3 in the plasma of patients with esophageal cancer, we also detected the EZH2, Bmi-1, P16, IMP-1 and Survivin expression changes of autoantibodies. Esophageal cancer and lung cancer antibody expression trends similar to P16 and Survivin autoantibodies The expression increased significantly (p0.001; p0.001), to analyze the tumor staging classification, P16 and Survivin autoantibodies in early esophageal carcinoma increased significantly as lung cancer. The subjects were ROC curve analysis, the specificity was 90% cases, esophageal cancer detection sensitivity were 26% and 15%.4 for better determine the specific tumor related antigens in the diagnosis of different tumors, in plasma of breast cancer patients, we also detected the EZH2, Bmi-1, P16, IMP-1 and Survivin antibody expression changes of.P16 and Survivin antibody expression in breast cancer was slightly increased, there was significant difference with healthy group (p=0.013; p=0.000), no significant difference in the stage of tumor in human subjects. ROC analysis indicated that the specificity of 90% cases of breast cancer detection sensitivity were 20% and 16%.5 in liver cancer, lung cancer, esophageal cancer and breast cancer clinical traditional marker AFP EGFR and HER2, for hierarchical phenotype analysis EZH2, Bmi-1, P16, IMP-1 and Survivin antibodies found in the expression of different phenotypes, HER2 and EGFR expression has nothing to do with the expression of autoantibodies. Interestingly, the expression of liver cancer patients are positively related to the expression of AFP and EZH2 and Bmi-1 autoantibodies. Conclusion: the expression of 1EZH2 Bmi-1 and autoantibodies in patients with primary hepatocellular carcinoma was significantly higher, while in other cancer cells did not change significantly, EZH2 and Bmi-1 antibodies may be potential biomarkers for liver cancer, has certain auxiliary effect on the diagnosis of hepatocellular carcinoma, and the expression of AFP had significant correlation. The combined detection can improve the detection sensitivity of hepatocellular carcinoma and the specificity of.2 P16 and Survivin in non small cell lung cancer antibody expression level was significantly higher in patients with lung cancer, including P16 and Survivin autoantibodies, the sensitivity and specificity of the best, can be used as lung Cancer diagnosis index, further optimization of P16 and Survivin.3 in esophageal carcinoma and autoantibody expression level of significant differences between the tumor group and the healthy people, but not more obvious in lung cancer, can be used as reference for.4Survivin diagnosis of esophageal cancer antibody expression in breast cancer was slightly elevated, while Survivin autoantibodies in lung and esophagus cancer also increased significantly, but in more than 3 kinds of tumor in the detection sensitivity of the differences, to further expand the sample to verify the specificity of.5 as markers of autoantibody detection in early diagnosis of tumor has certain auxiliary effect, has become a new method for the diagnosis of tumor potential.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R730.3
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本文编号：1552327