联合检测尿液中肿瘤标记物诊断膀胱癌的可行性研究

发布时间：2018-07-23 11:06

【摘要】：目的:全球范围内,每年发生约38万例膀胱恶性肿瘤,而在中国,膀胱癌是最多见的泌尿系肿瘤。膀胱癌的发病初期诊断和术后进行随访主要采用膀胱镜检查术,但它价钱昂贵且是一个侵入性的手段;尿细胞学检测方便,无创,但灵敏度不是很高,这些缺陷限制了膀胱癌的早期诊断和治疗。此外,膀胱癌的高复发率特点使得术后复发的监测尤为重要,但目前仍缺乏相应手段,这些问题使得一种新的检测手段成为需求。人的体液以及组织中具有一些特异性的肿瘤标记物(Tumormakers),它们可以用来特异性的检测某种恶性疾病,正因为它们对某种恶性肿瘤具有特异性,故而临床上常常应用肿瘤标记物来诊断特定肿瘤,并可以为肿瘤的治疗及预后进行评估,是以肿瘤标志物逐渐得到广泛的关注,成为当前热门的研究方向。本研究主要通过ELISA的方法检测尿液中NMP22、CD44、CK20、CEACAM1和BTA五种肿瘤标记物在膀胱癌患者尿液中的表达情况,并探讨上述五种标记物不同组合在膀胱癌诊断中的临床意义,为后续开发更为有效的能同时检测多种肿瘤标记物的试剂盒提供理论依据。方法:选取了2015年12月至2016年6月期间因怀疑膀胱肿瘤收治于天津医科大学第二医院的患者,经膀胱镜活检或TURBT术后病理诊断为膀胱癌的64名患者纳入本研究,设为肿瘤组;并选取20名非泌尿系肿瘤的患者作为对照组。应用酶联免疫吸附测定法(enzyme linked immunosorbent assay)分别检测两组患者尿液样本中NMP22、CD44、CK20、CEACAM1和BTA的表达水平,通过吖啶橙法(AO-F)行尿脱落细胞检查。应用SPSS19.0软件绘制受试者工作曲线(ROC),使用约登指数(Youden index)来计算5种肿瘤标记物的临界值(Cutoff),从而比较这五种肿瘤标记物以及不同组合联合诊断膀胱癌的准确性。应用秩和检验或费氏确切概率法分析五种肿瘤标记物与患者临床病理特征之间的关系。结果:1、NMP22、CD44、CK20、CEACAM1和BTA五种肿瘤标记物在肿瘤组的浓度显著高于对照组,差异具有统计学意义(P0.05)。2、NMP22、BTA、CK20、CD44、CEACAM1的曲线下面积(AUC)分别为0.752、0.820、0.773、0.859、0.724,由此可见这几种种标记物比较,NMP22检测膀胱癌的阳性率最高,达到84.3%,但特异度却很低,仅为55%。而尿脱落细胞AO-F法检测膀胱癌的特异度最高为100%,但它检测膀胱癌的阳性率却是6种肿瘤标记物中最低的,仅为60.9%。依据AUC比较,CD44的检测价值最大,当CD44取cutoff值为348.8pg/ml时,其敏感度可达82.8%,特异度为85%。3、尿脱落细胞AO-F法对于低分级尿路上皮癌的检测敏感度很低,仅为40%,而在高级别膀胱癌中的诊断敏感度为70.5%,两者有统计学差异(p=0.028)。尿AO-F法和CEACAM1检测膀胱癌的敏感度随肿瘤分期升高而逐渐增加,但其他各项指标却没有发现这种趋势。4、BTA、CD44、CK20三项联合检测时,敏感度为96.8%,特异度为80.0%。结论:1、应用ELISA法检测尿液中NMP22、BTA、CK20、CD44、CEACAM1这5种肿瘤标志物对膀胱癌的诊断具有一定的临床意义。2、应用1种标记物检测膀胱癌,CD44较其它4种标记物的价值最大3、CEACAM1对高分期膀胱癌的敏感度更高,尿脱落细胞AO-F检测对高分期、高分级膀胱癌的检测敏感度更高。4、CD44、CK20、BTA三者联合对膀胱癌的检测具有较高的临床意义。
[Abstract]:Objective: around 380 thousand cases of malignant bladder cancer occur annually worldwide, and bladder cancer is the most common urological tumor in China. Cystoscopy is mainly used in the early diagnosis and postoperative follow-up of bladder cancer, but it is expensive and an invasive hand; urine cytology is convenient and noninvasive, but the sensitivity is not It is very high, these limitations limit the early diagnosis and treatment of bladder cancer. In addition, the high recurrence rate of bladder cancer is particularly important for the monitoring of postoperative recurrence, but there is still a lack of corresponding means. These problems make a new detection method a demand. Human body fluid and group have some specific tumor markers (Tumorma KERS), they can be used to detect certain malignant diseases specifically. Because they are specific to a certain malignant tumor, they often use tumor markers to diagnose specific tumors, and can evaluate the treatment and prognosis of the tumor. It is gradually gaining extensive attention and becoming a hot research topic. The purpose of this study is to detect the expression of five tumor markers in urine of bladder cancer patients in urine by using ELISA method, and to explore the clinical significance of the different combinations of these five markers in the diagnosis of bladder cancer and to detect a variety of tumor markers at the same time for the subsequent development of the five kinds of markers. Methods: 64 patients who were admitted to Second Hospital Affiliated to Tianjin Medical University from December 2015 to June 2016 were enrolled in this study for bladder cancer with cystoscopy or TURBT, and 20 patients with non urinary tumors were selected as tumor groups. In the control group, the expression of NMP22, CD44, CK20, CEACAM1 and BTA in urine samples of two groups of two patients was detected by enzyme linked immunosorbent assay (linked immunosorbent assay), and the urine exfoliative cells were examined by acridine orange (AO-F). The working curve (ROC) was plotted with SPSS19.0 software, and the yonen index was used. The critical value of 5 tumor markers (Cutoff) was calculated to compare the accuracy of the five tumor markers and the combination of different combinations in the diagnosis of bladder cancer. The relationship between the five tumor markers and the clinicopathological features of the five tumor markers was analyzed by the rank sum test or the Fisher exact probability method. Results: 1, NMP22, CD44, CK20, CEACAM1, and BTA five tumor markers The concentration of the records in the tumor group was significantly higher than that in the control group. The difference was statistically significant (P0.05).2, NMP22, BTA, CK20, CD44, and CEACAM1 under the curves of AUC, respectively, 0.752,0.820,0.773,0.859,0.724. Thus, the positive rate of bladder cancer detected by NMP22 was the highest, reaching 84.3%, but the specificity was very low, only 55%.. The highest specificity of urinary exfoliative cell AO-F method for detecting bladder cancer was 100%, but the positive rate of bladder cancer detection was the lowest in 6 tumor markers. Only 60.9%. was compared with AUC, and CD44 was of the greatest value. When CD44 was 348.8pg/ml, its sensitivity was 82.8%, specificity was 85%.3, and urine exfoliative cells AO-F method. The detection sensitivity of low grade urothelial carcinoma was very low, only 40%, and the diagnostic sensitivity in advanced bladder cancer was 70.5%. There was a statistical difference between them (p=0.028). The sensitivity of urinary AO-F and CEACAM1 to bladder cancer increased with the increase of tumor stage, but the other indicators did not find the trend.4, BTA, CD44, CK20 three. The sensitivity of the joint examination was 96.8% and the specificity was 80.0%. conclusion: 1. The use of ELISA to detect NMP22 in urine, BTA, CK20, CD44, CEACAM1 has certain clinical significance for the diagnosis of bladder cancer, and the use of 1 markers for the detection of bladder cancer, the maximum value of CD44 compared with the other 4 markers is 3, CEACAM1 for high staging bladder cancer. The sensitivity of AO-F in urine exfoliative cells is higher than that of high staging, high grade bladder cancer, and.4, CD44, CK20, BTA three are of high clinical significance for the detection of bladder cancer.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14

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