BCM29和C3K5抗体检测膀胱癌细胞的实验研究

发布时间：2018-05-08 08:24

本文选题：循环肿瘤细胞 + 尿脱落细胞　；参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]探讨将巴氏染色作为对照,比较单克隆抗体BCM29和C3K5对膀胱癌细胞系细胞,膀胱癌患者尿脱落细胞、膀胱癌组织和循环肿瘤细胞的特异性和敏感性,初步评价其在膀胱尿路上皮癌的诊断和临床运用方面的运用价值。[方法](1)通过体外培养的膀胱尿路上皮癌细胞系EJ,确定易于观察的细胞系浓度,后以相对适浓度的膀胱癌细胞系EJ、T24和人胚肾细胞系293T分别用BCM29和C3K5抗体进行免疫组化染色和巴氏染色;(2)对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,分别用BCM29和C3K5抗体进行免疫组化染色和巴氏染色检测尿脱落细胞中的膀胱癌细胞,并统计出其检出率;(3)对20例膀胱尿路上皮癌患者肿瘤组织标本,以BCM29和C3K5抗体进行免疫组化染色检测两种抗体与肿瘤组织中癌细胞结合的特异性和敏感性;(4)对9例膀胱尿路上皮癌患者外周血通过红细胞裂解液裂解红细胞和CD45免疫磁珠去除白细胞阴性富集循环肿瘤细胞的方式,以BCM29抗体进行免疫组化染色和巴氏染色来检测循环肿瘤细胞。[结果](1)当细胞个数为0.5×106个时密度适中,易于观察;(2)两种抗体均可与膀胱尿路上皮癌细胞结合,与人胚肾细胞不结合;BCM29抗体浓度为1:105时染色效果佳;C3K5抗体浓度为1:2000时较佳。(3)BCM29和C3K5抗体均可与膀胱尿路上皮癌尿脱落细胞中的癌细胞结合,BCM29抗体对膀胱尿路上皮癌细胞敏感性为65.38%、特异性为94.44%,C3K5抗体敏感性为76.92%、特异性为61.11%,巴氏染色敏感性为34.62%、特异性为41.67%(敏感性:X2=10.28,P=0.006;特异性:X2=22.776,P=0.000),C3K5 抗体较易与白细胞结合。(4)两种抗体均可与膀胱癌组织中膀胱尿路上皮癌细胞结合;(5)在2例标本中检测出具有恶性特征和免疫组化阳性的疑似CTC,检出率为 22.22%。[结论](1).BCM29和C3K5抗体可与膀胱癌细胞系EJ、T24、膀胱癌组织、膀胱癌尿脱落细胞中的膀胱癌细胞结合,与人胚肾细胞293T和正常人尿路上皮细胞不结合;(2)BCM29和C3K5抗体对膀胱癌患者尿中癌细胞的特异性和敏感性均高于巴氏染色;(3)BCM29抗体对于膀胱癌细胞检测的特异性优于C3K5抗体;C3K5抗体对于膀胱癌细胞敏感性高于BCM29抗体;(4)BCM29和C3K5抗体在膀胱癌诊断方面有用较好的应用前景。
[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.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14

【参考文献】