發(fā)布時(shí)間：2018-05-08 08:24

  本文選題：循環(huán)腫瘤細(xì)胞 + 尿脫落細(xì)胞�。� 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]探討將巴氏染色作為對照,比較單克隆抗體BCM29和C3K5對膀胱癌細(xì)胞系細(xì)胞,膀胱癌患者尿脫落細(xì)胞、膀胱癌組織和循環(huán)腫瘤細(xì)胞的特異性和敏感性,初步評價(jià)其在膀胱尿路上皮癌的診斷和臨床運(yùn)用方面的運(yùn)用價(jià)值。[方法](1)通過體外培養(yǎng)的膀胱尿路上皮癌細(xì)胞系EJ,確定易于觀察的細(xì)胞 系濃度,后以相對適濃度的膀胱癌細(xì)胞系EJ、T24和人胚腎細(xì)胞系293T分別用BCM29和C3K5抗體進(jìn)行免疫組化染色和巴氏染色;(2)對26例膀胱尿路上皮癌患者尿液、36例非膀胱尿路上皮癌患者尿液和5例正常志愿者尿液,分別用BCM29和C3K5抗體進(jìn)行免疫組化染色和巴氏染色檢測尿脫落細(xì)胞中的膀胱癌細(xì)胞,并統(tǒng)計(jì)出其檢出率;(3)對20例膀胱尿路上皮癌患者腫瘤組織標(biāo)本,以BCM29和C3K5抗體進(jìn)行免疫組化染色檢測兩種抗體與腫瘤組織中癌細(xì)胞結(jié)合的特異性和敏感性;(4)對9例膀胱尿路上皮癌患者外周血通過紅細(xì)胞裂解液裂解紅細(xì)胞和CD45免疫磁珠去除白細(xì)胞陰性富集循環(huán)腫瘤細(xì)胞的方式,以BCM29抗體進(jìn)行免疫組化染色和巴氏染色來檢測循環(huán)腫瘤細(xì)胞。[結(jié)果](1)當(dāng)細(xì)胞個(gè)數(shù)為0.5×106個(gè)時(shí)密度適中,易于觀察;(2)兩種抗體均可與膀胱尿路上皮癌細(xì)胞結(jié)合,與人胚腎細(xì)胞不結(jié)合;BCM29抗體濃度為1:105時(shí)染色效果佳;C3K5抗體濃度為1:2000時(shí)較佳。(3)BCM29和C3K5抗體均可與膀胱尿路上皮癌尿脫落細(xì)胞中的癌細(xì)胞結(jié)合,BCM29抗體對膀胱尿路上皮癌細(xì)胞敏感性為65.38%、特異性為94.44%,C3K5抗體敏感性為76.92%、特異性為61.11%,巴氏染色敏感性為34.62%、特異性為41.67%(敏感性:X2=10.28,P=0.006;特異性:X2=22.776,P=0.000),C3K5 抗體較易與白細(xì)胞結(jié)合。(4)兩種抗體均可與膀胱癌組織中膀胱尿路上皮癌細(xì)胞結(jié)合;(5)在2例標(biāo)本中檢測出具有惡性特征和免疫組化陽性的疑似CTC,檢出率為 22.22%。[結(jié)論](1).BCM29和C3K5抗體可與膀胱癌細(xì)胞系EJ、T24、膀胱癌組織、膀胱癌尿脫落細(xì)胞中的膀胱癌細(xì)胞結(jié)合,與人胚腎細(xì)胞293T和正常人尿路上皮細(xì)胞不結(jié)合;(2)BCM29和C3K5抗體對膀胱癌患者尿中癌細(xì)胞的特異性和敏感性均高于巴氏染色;(3)BCM29抗體對于膀胱癌細(xì)胞檢測的特異性優(yōu)于C3K5抗體;C3K5抗體對于膀胱癌細(xì)胞敏感性高于BCM29抗體;(4)BCM29和C3K5抗體在膀胱癌診斷方面有用較好的應(yīng)用前景。
[Abstract]:[objective] to compare the specificity and sensitivity of monoclonal antibodies BCM29 and C3K5 to bladder cancer cell line, urinary exfoliative cells, bladder cancer tissue and circulating tumor cells. To evaluate its value in the diagnosis and clinical application of bladder urothelial carcinoma. [methods] the cell line of bladder urothelial carcinoma was cultured in vitro to determine the cell line concentration which was easy to observe. Then the bladder cancer cell line EJT24 and human embryonic kidney cell line 293T were used for immunohistochemical staining and pasteurized staining with BCM29 and C3K5 antibodies respectively. The urine samples of 26 patients with bladder urothelial carcinoma were divided into two groups: 36 cases of non-bladder urothelial carcinoma and 36 cases of non-bladder urothelial carcinoma. And 5 normal volunteers, BCM29 and C3K5 antibodies were used to detect bladder cancer cells in urine exfoliated cells by immunohistochemical staining and pasteurized staining respectively. BCM29 and C3K5 antibodies were used to detect the specificity and sensitivity of the two antibodies to cancer cells in tumor tissues. Nine patients with bladder urothelial carcinoma were treated with RBC and CD45 in peripheral blood. The way in which immunomagnetic beads remove leukocyte negative and enrich circulating tumor cells, Circulating tumor cells were detected by immunohistochemical staining and pasteurization with BCM29 antibody. [results] when the cell number was 0.5 脳 106, the cell density was moderate and it was easy to be observed.) both antibodies could bind to bladder urothelial carcinoma cells. When the antibody concentration of BCM29 with human embryonic kidney cells is 1: 105, the best staining effect is when the antibody concentration of C3K5 is 1: 2000. Both BCM29 and C3K5 antibodies can be used to bind BCM29 antibody in exfoliated cells of bladder urothelial carcinoma to bladder urothelial carcinoma. The cell sensitivity was 65.38, the specificity was 94.44 and the sensitivity of C3K5 antibody was 76.92, the specificity was 61.11, the sensitivity of pasteurization was 34.62 and the specificity was 41.67. The suspected CTC with malignant and immunohistochemical positive was detected in 2 cases of bladder urothelial carcinoma cells. The detection rate was 22.22%. [conclusion] the antibody to BCM29 and C3K5 can bind to bladder cancer cell line EJT24, bladder cancer tissue, bladder cancer exfoliated cells, bladder cancer cells. The specificity and sensitivity of BCM29 and C3K5 antibodies against human embryonic kidney cells 293T and normal human urinary tract epithelial cells were higher than those of pasteurian staining for detection of bladder cancer cells compared with C3K5. The sensitivity of anti-C _ 3K _ 5 antibody to bladder cancer cell is higher than that of BCM29 antibody, BCM29 and C3K5 antibodies in the diagnosis of bladder cancer.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.14

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本文編號：1860674