【摘要】：研究背景及目的：膀胱癌是威脅人類健康最常見的泌尿系統(tǒng)惡性腫瘤之一,其發(fā)病率及死亡率不斷上升。由于膀胱癌發(fā)病率的不斷增加以及膀胱癌的高復(fù)發(fā)率和多中心性,使其成為泌尿外科醫(yī)生及患者承受壓力最重的腫瘤之一。目前膀胱鏡檢查仍是膀胱癌診斷和隨訪的金標(biāo)準(zhǔn),但膀胱鏡為侵入性的檢查,給患者造成很大痛苦,不宜被患者所接受；同時細(xì)胞學(xué)檢查的敏感性低。目前還沒有非常理想的分子標(biāo)志物應(yīng)用于臨床。所以,尋找新的腫瘤標(biāo)志物以提高膀胱癌診斷的準(zhǔn)確性和及時性是一個迫切需要解決的問題。研究方法：首先,我們回顧性研究了北京協(xié)和醫(yī)院膀胱癌患者的臨床資料,收集了北京協(xié)和醫(yī)院有完整臨床病理和5年以上隨訪資料的患者,選擇膀胱癌患者及相應(yīng)病理切片共計(jì)140例,回顧性的研究了SNCG(synuclein-γ,神經(jīng)突觸核蛋白γ)、ARD1(arrest defective 1,N-乙�；D(zhuǎn)移酶調(diào)節(jié)亞基)及PRL-3 (phosphatase of regeneration liver-3,促肝再生磷酸酶3)在膀胱癌組織中的表達(dá)及其與膀胱癌臨床病理學(xué)特點(diǎn)和預(yù)后的關(guān)系；其次,我們選擇了其中在膀胱癌組織中表達(dá)最高的SNCG作為研究對象,進(jìn)一步探索研究了76例膀胱癌患者血清及尿液SNCG蛋白表達(dá),并分析了SNCG的表達(dá)及分泌方式,最后,為研究尿液SNCG檢測的臨床價值,我們進(jìn)行了大規(guī)模的,多中心的隊(duì)列研究,以驗(yàn)證尿液SNCG作為膀胱癌診斷和術(shù)后監(jiān)測工具的價值,并與NMP22進(jìn)行了對比研究。研究結(jié)果：140例膀胱癌組織通過免疫組化研究發(fā)現(xiàn)SNCG蛋白的陽性表達(dá)率為90.7%(127/140)；ARD1蛋白的陽性表達(dá)率為80.7%(113/140)；PRL-3蛋白的陽性表達(dá)率為17.1%(24/140)。膀胱癌組織中SNCG蛋白的表達(dá)方式較為復(fù)雜,有細(xì)胞核陽性表達(dá)、也有彌漫細(xì)胞漿和細(xì)胞膜表達(dá),總體陽性率較高,并提示膀胱癌細(xì)胞能自主分泌SNCG蛋白。前期試驗(yàn)證實(shí)SNCG蛋白可以從MCF7-SNCG細(xì)胞中分泌到細(xì)胞外的培養(yǎng)上清中,證明SNCG是一種分泌性蛋白,且為腫瘤細(xì)胞特異表達(dá)的蛋白,膀胱癌細(xì)胞既可以分泌SNCG入血,也可以直接分泌到腫瘤周圍的尿液中。膀胱癌患者血清中的SNCG水平明顯高于正常人,而同一患者尿液中SNCG濃度明顯高于血清中的水平。同時我們建立的雙抗夾心ELISA法檢測SNCG水平具有很好的特異性、穩(wěn)定性和可靠性。大規(guī)模多中心的隊(duì)列研究證明了膀胱癌患者尿SNCG濃度顯著高于年齡和性別相匹配的健康對照組。在測試隊(duì)列中膀胱癌組SNCG診斷試驗(yàn)的ROC曲線下面積(area under the ROC curve,AUC)為0.903±0.019(95%可信區(qū)間為0.867至0.940)；在驗(yàn)證隊(duì)列中的ROC曲線下面積為0.929±0.015(95%可信區(qū)間0.901至0.958)。Youden指數(shù)越大,說明診斷準(zhǔn)確度越高。根據(jù)這一原則,最終確認(rèn)1.874納克／毫升作為最佳診斷界值,在測試隊(duì)列得到的敏感度及特異度分別為68.4%和97.4%；在驗(yàn)證隊(duì)列中得到的敏感度和特異度分別為62.4%和97.8%；。此外,我們發(fā)現(xiàn),膀胱癌患者手術(shù)后尿液SNCG下降,術(shù)后復(fù)查時SNCG水平在復(fù)發(fā)患者(17.18±44.19 ng/mL)比無復(fù)發(fā)患者(2.81±11.95 ng/mL, P=0.001)要高。在一個小的隊(duì)列研究中,膀胱癌診斷試驗(yàn)結(jié)果發(fā)現(xiàn)SNCG敏感性較NMP22稍低(46.2%對56.4%,P=0.344)但特異性較NMP22高(74.5%對51.0%,P=0.008), SNCG診斷效能優(yōu)于NMP22,且不受血尿因素影響。研究結(jié)論：SNCG作為一種分泌性蛋白,不僅在膀胱癌組織中有極高的表達(dá),在膀胱癌患者的尿液中也有較高的表達(dá),尿液SNCG檢測具有鑒別膀胱癌患者與健康人及泌尿系統(tǒng)其他良性病變的能力,且尿液SNCG檢測并不受血尿的影響。不僅具有診斷早期膀胱癌的價值,對膀胱癌術(shù)后復(fù)發(fā)也有很好的監(jiān)測作用。SNCG表達(dá)與癌細(xì)胞增殖、轉(zhuǎn)移和藥物抗性有關(guān)。檢測癌組織SNCG水平可預(yù)測腫瘤患者的預(yù)后；抑制SNCG活性可增加對化療藥物的敏感性,因此SNCG有望成為膀胱癌監(jiān)測的分子標(biāo)志物及治療的潛在靶點(diǎn)。
[Abstract]:The background and purpose of the study are that bladder cancer is one of the most common malignant tumor of the urinary system, and its morbidity and mortality are increasing. As a result of the increasing incidence of bladder cancer and the high recurrence rate and multi-center of bladder cancer, it is one of the most stressful tumors in urological surgeons and patients. At present, cystoscopy is still a gold standard for the diagnosis and follow-up of bladder cancer, but the bladder mirror is an invasive examination, which causes a great deal of pain to the patient and is not suitable to be accepted by the patient; at the same time, the sensitivity of the cytological examination is low. There is currently no very ideal molecular marker for clinical use. Therefore, finding new tumor markers to improve the accuracy and timeliness of bladder cancer diagnosis is an urgent problem. Methods: First of all, we retrospectively analyzed the clinical data of the patients with bladder cancer in Peking Union and Hospital, and collected a total of 140 patients with complete clinical pathology and more than 5 years of follow-up data in Peking Union Hospital, and 140 cases of bladder cancer patients and corresponding pathological sections were selected. In this paper, the expression of SNCG (synclinin-1, neurosynaptic nucleoprotein), ARD1 (arrestt 1, N-glycosyltransferase-regulating subunit) and PRL-3 (phosphatase of regrowth liver-3, prohepatic regeneration phosphatase 3) in the bladder cancer tissue and its relationship with the clinicopathological features and prognosis of bladder cancer were studied retrospectively. The expression of SNCG in serum and urine of 76 cases of bladder cancer was further explored, and the expression and secretion of SNCG were also analyzed. We conducted a large-scale, multicenter cohort study to verify the value of urine SNCG as a tool for bladder cancer diagnosis and post-operation monitoring and a comparative study with NMP22. Results: The positive expression rate of SNCG was 90.7% (127/140), the positive expression rate of ARD1 protein was 80.7% (113/140), and the positive expression rate of PRL-3 was 17.1% (24/140). The expression of SNCG in bladder cancer is more complex, with the positive expression of the cell nucleus, and the expression of diffuse cell and cell membrane, the overall positive rate is high, and it is suggested that the bladder cancer cell can secrete SNCG protein autonomously. The early test confirms that the SNCG protein can be secreted into the culture supernatant from the MCF7-SNCG cell to the outside of the cell to prove that the SNCG is a secreted protein and is a protein that is specifically expressed for the tumor cells, and the bladder cancer cell can secrete SNCG blood into the blood, and can be directly secreted into the urine around the tumor. The level of SNCG in the serum of patients with bladder cancer was significantly higher than that of the normal, while the concentration of SNCG in the urine of the same patient was significantly higher than that in the serum. At the same time, we established the double anti-sandwich ELISA method to detect the level of SNCG with good specificity, stability and reliability. The large-scale multi-center cohort study demonstrated that the urinary SNCG concentration in bladder cancer patients was significantly higher than that of the age and gender-matched healthy controls. The area under the ROC curve for the SNCG diagnostic test in the test cohort was 0.903-0.019 (95% confidence interval 0.867 to 0.940); the area under the ROC curve in the validation cohort was 0.929-0.015 (95% confidence interval 0.901 to 0.958). The greater the Eden index, the higher the diagnostic accuracy. The sensitivity and specificity in the test cohort were 68.4% and 97.4%, respectively, according to this principle. The sensitivity and specificity were 62.4% and 97.8%, respectively. In addition, we found that the urine SNCG decreased after the operation of the bladder cancer patients, and the SNCG level was higher in the patients with the recurrence (17.18, 44.19 ng/ mL) than in the non-recurrent patients (2.81, 11.95 ng/ mL, P = 0.001). In a small cohort study, the results of the diagnosis of bladder cancer found that the sensitivity of SNCG was slightly lower than that of NMP22 (46.2% vs 56.4%, P = 0.344), but the specificity was higher (74.5% vs 51.0%, P = 0.008), and the diagnostic efficacy of SNCG was superior to NMP22 and not affected by hematuria. Conclusion: SNCG is a kind of secretory protein, not only has very high expression in bladder cancer tissues, but also has high expression in the urine of bladder cancer patients. The detection of SNCG in urine has the ability to identify the bladder cancer patients and other benign diseases of the healthy people and the urinary system. And the urine SNCG is not affected by hematuria. Not only has the value of diagnosing early bladder cancer, but also has good monitoring effect on postoperative recurrence of bladder cancer. The expression of SNCG is related to the proliferation, metastasis and drug resistance of cancer cells. Detecting the level of SNCG in the cancer tissue can predict the prognosis of a tumor patient, and the inhibition of the SNCG activity can increase the sensitivity to the chemotherapy drug, so that the SNCG is expected to be a molecular marker for bladder cancer monitoring and a potential target for treatment.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R737.14

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 褚亮;萬圣云;周少波;劉會春;高涌;;CA19-9、CA242及CEA聯(lián)合檢測在良惡性梗阻性黃疸鑒別診斷中的應(yīng)用[J];蚌埠醫(yī)學(xué)院學(xué)報;2011年10期

2 謝瑋;蘇亞輝;劉彩云;壽成超;;抗synuclein-γ單克隆抗體識別的抗原表位區(qū)域的確定[J];北京大學(xué)學(xué)報(醫(yī)學(xué)版);2009年03期

3 江濱;張紅英;譚妍妍;陳艷妮;徐斐;丁義江;;神經(jīng)核蛋白γ(γ-syn)在直腸癌組織中表達(dá)的研究[J];結(jié)直腸肛門外科;2010年02期

4 徐衛(wèi)東;趙建國;殷輝;焦鋼;王曉燕;;原發(fā)性骨肉瘤中γ-synuclein的表達(dá)[J];第二軍醫(yī)大學(xué)學(xué)報;2007年04期

5 Gareth Morris-Stiff;Mary Teli;Nicky Jardine;Malcolm CA Puntis;;CA19-9 antigen levels can distinguish between benign and malignant pancreaticobiliary disease[J];Hepatobiliary & Pancreatic Diseases International;2009年06期

6 何幫順;王書奎;;乳腺癌特異性基因1的生物學(xué)特性及與惡性腫瘤的關(guān)系[J];國際檢驗(yàn)醫(yī)學(xué)雜志;2006年04期

7 張玲,李兆申;蛋白質(zhì)組學(xué)在胰腺癌早期診斷中的研究進(jìn)展[J];國外醫(yī)學(xué)(消化系疾病分冊);2005年05期

8 郭劍平;孟麟;壽成超;;γ-突觸核蛋白與腫瘤的關(guān)系[J];國際腫瘤學(xué)雜志;2006年11期

9 鄒靖;范余娟;徐紅;蒙亞晴;范江濤;徐文生;;γ-synuclein在子宮內(nèi)膜癌中的表達(dá)及其與預(yù)后的關(guān)系[J];廣西醫(yī)學(xué);2012年02期

10 王天昱;陳曉鵬;李學(xué)松;賈元歆;成俊;張建華;蔡林;張爭;龔侃;何志嵩;周利群;;前列腺特異性抗原和Gleason評分對前列腺癌患者核素骨掃描結(jié)果的預(yù)測價值[J];北京大學(xué)學(xué)報(醫(yī)學(xué)版);2012年04期

相關(guān)會議論文 前8條

1 吳沛宏;黃職妹;;瘤區(qū)淋巴結(jié)的新治療策略:陽性清除之,陰性保留之,可疑觀察之[A];第九屆中國腫瘤微創(chuàng)治療學(xué)術(shù)大會論文集[C];2013年

2 鄭偉慧;毛偉敏;夏李明;;喉返神經(jīng)旁淋巴結(jié)清掃對胸段食管癌TNM分期影響[A];2013華東胸部腫瘤論壇暨第六屆浙江省胸部腫瘤論壇論文集[C];2013年

3 鄧勇;張煒飛;張成斌;林金明;;液相色譜串聯(lián)質(zhì)譜法定量檢測前列腺癌細(xì)胞肌氨酸代謝[A];中國化學(xué)會第29屆學(xué)術(shù)年會摘要集——第38分會：質(zhì)譜分析[C];2014年

4 支修益;;亞肺葉切除治療早期肺癌[A];中國腫瘤內(nèi)科進(jìn)展 中國腫瘤醫(yī)師教育（2014）[C];2014年

5 張蕾;王崇薇;史天陸;張圣雨;孫言才;姜玲;;2004-2013年我院腫瘤患者嚴(yán)重藥品不良反應(yīng)分析[A];第十三屆全國青年藥師成才之路論壇暨抗腫瘤藥物合理應(yīng)用與臨床藥學(xué)實(shí)踐國家級繼教會議論文集[C];2014年

6 鄧勇;張煒飛;張成斌;林金明;;姜黃素對前列腺癌細(xì)胞肌氨酸代謝和AR/TMPRSS2-ERG信號通路的影響[A];中國化學(xué)會首屆全國質(zhì)譜分析學(xué)術(shù)研討會會議論文集[C];2014年

7 謝蕓;王文杰;;肺癌患者手術(shù)后心律失常原因及護(hù)理對策分析[A];中國轉(zhuǎn)化醫(yī)學(xué)和整合醫(yī)學(xué)研討會（廣州站）論文綜合刊[C];2015年

8 劉瑞;何姝霖;鄭紅剛;李衛(wèi)東;花寶金;齊鑫;裴迎霞;張蕓;;肺瘤平膏及其聯(lián)合塞來昔布干預(yù)時間對Lewis肺癌小鼠不同腫瘤微環(huán)境中COX-2表達(dá)的影響[A];第一屆青年中西醫(yī)結(jié)合腫瘤學(xué)術(shù)論壇論文集[C];2015年

相關(guān)博士學(xué)位論文 前10條

1 陳志康;TMPRSS4表達(dá)調(diào)控與胃癌侵襲轉(zhuǎn)移的關(guān)系研究[D];中南大學(xué);2011年

2 申培紅;siRNA對乳腺癌特異性基因表達(dá)影響及與S100A4、ETS1、VEGF、CD105基因表達(dá)關(guān)系的研究[D];鄭州大學(xué);2011年

3 王媛媛;大腸癌干細(xì)胞基因表達(dá)譜特點(diǎn)及其信號通路的篩選鑒定[D];南方醫(yī)科大學(xué);2011年

4 韓為農(nóng);鼻咽低分化鱗狀細(xì)胞癌的分子分類[D];中南大學(xué);2003年

5 孫立峰;SNC19/ST14基因的表達(dá)對大腸癌細(xì)胞生物學(xué)特性及其轉(zhuǎn)移相關(guān)基因表達(dá)的影響[D];浙江大學(xué);2004年

6 劉巍;乳腺癌maspin、BCSG1表達(dá)情況及新輔助化療對其表達(dá)影響的研究[D];河北醫(yī)科大學(xué);2005年

7 周曉波;人乳頭瘤病毒（HPV）在中國食管癌中的檢測以及HPV16 E7作用機(jī)制的研究[D];中國協(xié)和醫(yī)科大學(xué);2003年

8 張玲;胰腺癌中ELA3B和NPTX2基因甲基化狀態(tài)的檢測及NPTX2基因的功能研究[D];第二軍醫(yī)大學(xué);2008年

9 顧玉梅;基因組CpG島甲基化譜的改變與腫瘤發(fā)生和轉(zhuǎn)移的關(guān)系[D];南京醫(yī)科大學(xué);2008年

10 郭春光;結(jié)直腸癌肝轉(zhuǎn)移相關(guān)分子標(biāo)志物的篩選與鑒定[D];中國協(xié)和醫(yī)科大學(xué);2008年

，

本文編號：2468134