本文關(guān)鍵詞： 自身抗體 診斷標(biāo)志物 肺癌 食管癌 乳腺癌 肝癌 腫瘤相關(guān)抗原　出處：《吉林大學(xué)》2016年博士論文　論文類型：學(xué)位論文

【摘要】：目的:中國是世界人口大國也是癌癥高發(fā)的國家,由于腫瘤多起病隱匿、早期表現(xiàn)不明顯,導(dǎo)致多數(shù)腫瘤患者在發(fā)現(xiàn)時就已經(jīng)處于中晚期階段,治療效果不理想患者生活質(zhì)量大大降低。所以腫瘤的早期診斷、早治療對腫瘤患者來說十分重要。然而目前腫瘤的診斷主要依賴于影像學(xué)和細(xì)胞學(xué)檢查,診斷能力有限,目前亟待出現(xiàn)一種能更早期診斷腫瘤的手段。自從Landsteier提出機(jī)體內(nèi)存在自身抗體(autoantibody)以來,人們逐漸對自身抗體有了深刻的認(rèn)識。在長期腫瘤發(fā)病機(jī)制研究過程中,學(xué)者們認(rèn)為基因突變是最基本的觸發(fā)環(huán)節(jié)或始動因素,這些異常表達(dá)的蛋白就成為了腫瘤抗原。相對于濃度較低的腫瘤抗原,自身抗體的檢測具有更高的敏感性。因此我們檢測高發(fā)腫瘤中自身抗體的表達(dá)變化,以期尋找到可用于腫瘤輔助診斷的新型腫瘤標(biāo)志物。方法:通過查閱文獻(xiàn)和課題組前期工作基礎(chǔ),本研究中納入5種目標(biāo)蛋白,分別是m RNA的結(jié)合蛋白IMP-1、凋亡抑制基因蛋白Survivin、腫瘤抑制因子P16、轉(zhuǎn)錄調(diào)節(jié)蛋白EZH2和原癌基因蛋白Bmi-1。利用免疫學(xué)分析軟件和表位數(shù)據(jù)庫,預(yù)測、設(shè)計并合成5種蛋白的多肽抗原片段,應(yīng)用經(jīng)過我們優(yōu)化改良的ELISA檢測方法,包被多肽抗原,根據(jù)免疫學(xué)抗原抗體結(jié)合原理檢測血漿中自身抗體水平。研究共納入未經(jīng)任何抗癌治療的首次診斷為惡性腫瘤的肝癌患者、食管癌患者、肺癌患者、乳腺癌患者和健康人群各200例。設(shè)置質(zhì)控血衡量檢測的穩(wěn)定性,根據(jù)酶標(biāo)儀測得的樣本OD和質(zhì)控OD計算相對結(jié)合指數(shù),降低非特異結(jié)合和檢測誤差。應(yīng)用SPSS21.0進(jìn)行數(shù)據(jù)統(tǒng)計和分析,比較自身抗體水平在不同腫瘤、不同分期中的表達(dá)變化,分析其在不同腫瘤中表達(dá)特點(diǎn)的同時,探討其在腫瘤診斷中的作用。結(jié)果:1肝癌患者血漿5種腫瘤相關(guān)抗原自身抗體檢測中,EZH2和Bmi-1自身抗體水平出現(xiàn)顯著升高(p0.001;p0.001),進(jìn)一步按腫瘤分期分析發(fā)現(xiàn),早期患者EZH2和Bmi-1自身抗體水平高于晚期患者。受試人群ROC曲線分析發(fā)現(xiàn),EZH2和Bmi-1自身抗體檢測在肝癌診斷中具有較高的應(yīng)用價值,特異度90%情況下,肝癌檢測敏感度分別為22%和32%。2在非小細(xì)胞肺癌患者血漿中,同樣檢測了EZH2、Bmi-1、P16、IMP-1和Survivin自身抗體表達(dá)變化。其中P16和Survivin自身抗體表達(dá)出現(xiàn)顯著升高(p0.001;p0.001),雖然IMP-1亦出現(xiàn)升高趨勢,但沒有統(tǒng)計學(xué)差異。以腫瘤分期分層分析中,P16和Survivin自身抗體在肺癌早期升高更明顯。受試人群ROC曲線分析發(fā)現(xiàn),P16和Survivin自身抗體在肺癌診斷中具有很好的臨床價值,特異度90%情況下其敏感度分別為32%和20%。3食管癌患者血漿中,我們同樣檢測了EZH2、Bmi-1、P16、IMP-1和Survivin自身抗體表達(dá)變化。食管癌和肺癌中自身抗體表達(dá)變化趨勢相似,P16和Survivin自身抗體表達(dá)出現(xiàn)顯著升高(p0.001;p0.001),以腫瘤分期分層分析中,P16和Survivin自身抗體在食管癌早期升高不如肺癌明顯。受試人群ROC曲線分析發(fā)現(xiàn),特異度90%情況下,食管癌檢出的敏感度分別為26%和15%。4為更好判斷腫瘤相關(guān)抗原自身抗體在不同腫瘤診斷中的特異性,在乳腺癌患者血漿中,我們同樣檢測了EZH2、Bmi-1、P16、IMP-1和Survivin自身抗體表達(dá)變化。P16和Survivin自身抗體表達(dá)在乳腺癌中略有升高,與健康組存在統(tǒng)計學(xué)差異(p=0.013;p=0.000),在腫瘤分期中沒有明顯的差異。受試人群ROC曲線分析發(fā)現(xiàn),特異度90%情況下,乳腺癌檢出的敏感度分別為20%和16%。5以肝癌、肺癌、食管癌和乳腺癌的臨床傳統(tǒng)標(biāo)志物AFP、EGFR和HER2為分層表型,分析EZH2、Bmi-1、P16、IMP-1和Survivin自身抗體在不同表型表達(dá)變化發(fā)現(xiàn),HER2和EGFR過表達(dá)與自身抗體表達(dá)無關(guān)。有趣的是,肝癌患者中AFP的表達(dá)與EZH2和Bmi-1自身抗體的表達(dá)呈現(xiàn)正相關(guān)。結(jié)論:1EZH2和Bmi-1自身抗體在肝癌患者中表達(dá)水平明顯升高,而在其他腫瘤中表達(dá)變化并不明顯,EZH2和Bmi-1自身抗體可能是肝癌的潛在生物學(xué)標(biāo)志物,對肝癌診斷有一定輔助作用,且與AFP的表達(dá)有顯著相關(guān)性。其聯(lián)合檢測可有效提高肝癌檢出的敏感度和特異度。2非小細(xì)胞肺癌中P16和Survivin自身抗體表達(dá)水平在肺癌患者中明顯升高,其中P16和Survivin自身抗體的敏感度和特異度最好,可以作為肺癌診斷指標(biāo),有待進(jìn)一步優(yōu)化。3食管癌中P16和Survivin自身抗體表達(dá)水平在腫瘤組和健康人群之間存在顯著差異,但不如肺癌中更明顯,可以作為食管癌輔助診斷的參考指標(biāo)。4Survivin自身抗體表達(dá)在乳腺癌中略有升高,雖然Survivin自身抗體在肺癌和食管癌也均明顯升高,但在以上3種腫瘤中其檢測的敏感度存在差異,需進(jìn)一步擴(kuò)大樣本來驗證其作為標(biāo)志物的特異性。5自身抗體檢測在腫瘤早期診斷中具有一定輔助作用,有成為腫瘤診斷新方法的潛能。
[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.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R730.3
，

本文編號：1552327