BLCA-1在肌层浸润性膀胱癌患者血清、尿液中的表达

发布时间：2017-12-28 01:31

本文关键词：BLCA-1在肌层浸润性膀胱癌患者血清、尿液中的表达　出处：《承德医学院》2016年硕士论文　论文类型：学位论文

【摘要】：在我国,膀胱癌是最常见的泌尿系恶性肿瘤之一,其逐年上升的发病率和高复发率给患者和社会带来了沉重的经济和医疗负担。目前诊断膀胱肿瘤的金标准依然是膀胱镜检查加组织活检,但临床中这种有创性操作具有潜在并发症,不适用于大规模人群筛查,而传统的尿脱落细胞学检查因其敏感性较低,同样不适用于膀胱癌的早期诊断、术后随访。因此寻找方便快捷敏感特异的肿瘤标志物成为了科研工作者们的研究热点。在肿瘤发生发展中,会释放以酶、激素、抗原等形式存在于患者机体环境中的肿瘤标记物。据研究,膀胱癌特异性核基质蛋白(Bladder cancer specific nuclear matrix proteins,BLCAs)家族特异表达于膀胱肿瘤组织,其中BLCA-1显示出了较高的敏感性和特异性。本研究应用酶联免疫吸附实验(Enzyme linked immunosorbent assay,ELISA),对受试人群血清与尿液中的BLCA-1和膀胱肿瘤相关抗原(Bladder tumor antigen,BTA)进行定量检测,探讨BLCA-1与膀胱癌生物学特性的内在联系,并与FDA认可的肿瘤标志物BTA检测结果及本课题组王晓鹏等人关于BLCA-4的检测结果相对比,为开发在膀胱癌筛查、诊断及术后随访中具有高度特异性、敏感性的肿瘤标记物提供实验依据。目的:通过实验观测BLCA-1在受试人群血清、尿液中的表达水平,探讨BLCA-1在膀胱癌早期筛查、诊断及术后监测中的应用价值。方法:受试人群分为实验组、对照组,选取我院2014年3月~2015年3月22例病理结果确认为肌层浸润性膀胱尿路上皮癌患者作为实验组,其中男16例,女6例,年龄范围45~90岁,平均年龄64±11岁。根据WHO2004分级法,乳头状尿路上皮癌低分级:5例,高分级:17例。根据2009 TNM分期标准,T2:16例,T3:5例,T4:1例。对照组包括健康体检者和良性前列腺增生患者共44例,对照组A:选自健康体检者22例,其中男10例,女12例,年龄范围21~66岁,平均年龄41±14岁。对照组B:良性前列腺增生症患者22例,年龄范围52~81岁,平均年龄64±7岁。三组受试人群一般情况无差异,可以比较。采用elisa法定量检测22例肌层浸润性膀胱癌患者、44例对照组患者血清与尿液blca-1和上述人群尿液中bta的表达水平。检测blca-4在肌层浸润性膀胱癌中的表达已由本课题组的王晓鹏等人完成,实验对象与试验方法与本实验一致,具有可比性。应用统计学分析实验数据。非正态性计量资料均用中位数加减四分位间距(m±qd)表示,非正态性计量资料两两比较采用wilcoxon秩和检验,三组间差异进行kruskal-wallish检验,多组间两两比较采用nemenyi检验;blca-1与bta之间比较采用fisher确切概率法检验;计数资料用百分比表示,所有资料均应用spss19.0统计软件进行处理,以p0.05为差异有统计学意义。结果:实验组、对照组a、对照组b受试者尿液blca-1含量的中位数为1.479ng/ml、0.435ng/ml和0.709ng/ml,实验组患者blca-1表达水平显著高于其它两组(p0.01),对照组b患者blca-1表达水平较对照组a升高,差异有统计学意义(p0.05)。当cut-off值取0.946ng/ml时,尿blca-1敏感性、特异性最佳,分别为77%(17/22)和93%(41/44)。实验组、对照组a、对照组b受试者血清blca-1含量的中位数分别为6.621ng/ml、3.522ng/ml和5.611ng/ml,实验组与对照组b血清blca-1表达水平差异无统计学意义(p0.05),明显高于对照组a(p0.01)。当cut-off值取4.537ng/ml时,血blca-1敏感性、特异性最佳,分别为86%(19/22)和56%(26/44)。实验组、对照组a和对照组b尿液中bta含量中位数分别为214.494ng/ml、9.384ng/ml和121.982ng/ml,实验组数据高于两对照组(p0.05),三组数据两两比较差异均有统计学意义(p0.05)。当cut-off值取81.01ng/ml时,尿bta敏感性、特异性最佳,分别为77%(17/22)和64%(28/43)。实验组尿液blca-1表达水平与患者年龄、性别、肌层浸润性膀胱癌不同分期、分级无相关性,差异无统计学意义(p0.05);实验组血清blca-1水平在膀胱癌患者高年龄(≥60岁)和高分级中表达较高(p0.05)。应用fisher确切概率法比较elisa检测患者尿液blca-1和bta在肌层浸润性膀胱癌患者诊断中的敏感性和特异性,结果显示:两者敏感性相近(p0.05),尿blca-1特异性显著高于尿bta(p0.05)。同时,尿BLCA-1检测低级别肌层浸润性膀胱癌患者敏感性显著高于尿BTA(P0.05),分别为80%(4/5)、20%(1/5),特异性差异无统计学意义(P0.05),分别为88%(15/17)、94%(16/17)。ELISA检测尿液BLCA-1诊断膀胱癌的准确度、阳性预测值、阴性预测值和Youden指数均高于尿BTA。本课题组王晓鹏等人实验显示ELISA法检测尿中BLCA-4诊断膀胱癌敏感性为91%(22/24),特异性为100%,但血清BLCA-4的表达无诊断价值。应用Fisher精确概率法分析,尿BLCA-4的敏感性、特异性与本实验尿BLCA-1结果差异无统计学意义(P0.05)。结论:1、当cut-off值取0.946ng/ml,检测尿液BLCA-1诊断膀胱癌敏感性特异性最佳,但其表达水平与肌层浸润性膀胱癌不同分级、分期未显示出相关性。2、当cut-off值取4.537ng/ml时,检测血清BLCA-1诊断膀胱癌敏感性特异性最佳。患者血清中BLCA-1水平显示出了与肌层浸润性膀胱癌患者年龄和肿瘤分级的相关性,具体机制尚不明确,还需进一步研究。3、与具有代表性的膀胱肿瘤标志物BTA比较,尿BLCA-1显示出了较高特异性(93%vs 64%)。检测尿BLCA-1诊断膀胱癌阳性预测值、阴性预测值、Youden指数及准确性均高于BTA。4、作为新一代膀胱肿瘤标志物,竞争性ELISA法检测BLCA-1诊断膀胱癌简便无创,具有潜在临床价值。
[Abstract]:Bladder cancer is one of the most common urological malignancies in China. Its yearly rising incidence and high recurrence rate bring heavy economic and medical burden to patients and society. The current gold standard for diagnosis of bladder cancer is still the cystoscopy and biopsy, but the clinical invasive operation with potential complications, not suitable for large-scale screening, and conventional cytology due to its low sensitivity, early diagnosis and postoperative follow-up were also not suitable for bladder cancer. Therefore, it has become a hot spot for researchers to find the convenient and sensitive and specific tumor markers. In the development of tumor, the tumor markers that exist in the environment of the patient are released in the form of enzymes, hormones, antigens and other forms. According to the research, the Bladder cancer specific nuclear matrix proteins (BLCAs) family is specifically expressed in bladder tumor tissues, and BLCA-1 shows high sensitivity and specificity. The research and application of enzyme linked immunosorbent assay (Enzyme linked immunosorbent assay, ELISA), on the subject of BLCA-1 and bladder tumor associated antigens in serum and urine in the crowd (Bladder tumor, antigen, BTA) were detected, the relation of BLCA-1 and the biological characteristics of bladder cancer, and the detection results of BTA markers and the research group Wang Xiaopeng et al. Detection of BLCA-4 on the results compared with the FDA approved for the development of tumor in bladder cancer screening, diagnosis and postoperative follow-up with high specificity and sensitivity of tumor markers to provide experimental basis for. Objective: To observe the expression level of BLCA-1 in the serum and urine of subjects, and to explore the value of BLCA-1 in early screening, diagnosis and postoperative monitoring of bladder cancer. Methods: the subjects were divided into experimental group and control group. 22 cases of myometrial invasive bladder urothelial carcinoma confirmed by pathology in our hospital in March March 2014 ~2015 were selected as the experimental group, including 16 males and 6 females, with an age range of 45~90 years, with an average age of 64 + 11 years. According to the WHO2004 classification, the low grade of papillary urothelial carcinoma: 5 cases, high grade: 17 cases. According to the 2009 TNM staging criteria, T2:16 cases, T3:5 cases, and T4:1 cases. The control group consisted of 44 healthy subjects and 22 patients with benign prostatic hyperplasia. In the control group, A: was selected from 22 healthy subjects, including 10 males and 12 females. The age range was 21~66 years old, with an average age of 41 + 14 years. In the control group, there were 22 cases of B: benign prostatic hyperplasia (BPH), the age range of 52~81 years, the average age of 64 + 7 years. The general situation of the three groups was not different, and could be compared. The levels of serum and urine blca-1 and the expression level of BTA in urine of 22 cases of muscle invasive bladder cancer and 44 cases of control group were detected by ELISA. The detection of BLCA-4 expression in muscle invasive bladder cancer has been completed by Wang Xiaopeng, our research group. The experimental objects and experimental methods are consistent with this experiment. Statistical analysis of experimental data. The non normality of measurement data uses the median plus or minus four points spacing (M + QD) said that the non normality of measurement data of 22 compared with the Wilcoxon rank sum test, the difference between the three groups by kruskal-wallish test, multiple comparison between the 22 groups using nemenyi test; between blca-1 and BTA were compared by Fisher exact test; count data expressed as a percentage, all the data are processed by spss19.0 statistical software, using P0.05 as the difference was statistically significant. Results: the experimental group and the control group, the control group a median B subjects urine blca-1 content for 1.479ng/ml, 0.435ng/ml and 0.709ng/ml, the expression level of blca-1 in experimental group were significantly higher than the other two groups (P0.01), the control group B expression level of blca-1 in patients with a increased compared with the control group, the difference was statistically significant (P0.05). When the cut-off value was 0.946ng/ml, the sensitivity and specificity of urine blca-1 were the best, 77% (17/22) and 93% (41/44), respectively. The median level of serum blca-1 in experimental group, control group A and control group B was 6.621ng/ml, 3.522ng/ml and 5.611ng/ml, respectively. There was no significant difference in serum blca-1 expression level between experimental group and control group (P0.05), which was significantly higher than that of control group (P0.01). When the cut-off value was 4.537ng/ml, the sensitivity and specificity of blood blca-1 were the best, 86% (19/22) and 56% (26/44), respectively. The median BTA content of B urine in experimental group and control group A and control group were 214.494ng/ml, 9.384ng/ml and 121.982ng/ml, respectively. The data in experimental group were higher than those in two control group (P0.05), and the difference between three groups was statistically significant (P0.05). The difference between 22 groups was statistically significant (P0.05). When the cut-off value was 81.01ng/ml, the sensitivity and specificity of urine BTA were the best, 77% (17/22) and 64% (28/43), respectively. The experimental group of urine blca-1 expression and patient age, gender, muscle invasive bladder cancer in different stage and grade of no correlation, there was no significant difference between the experimental group (P0.05); the level of serum blca-1 in aged patients with bladder cancer (over 60 years) and high grade high expression (P0.05). The sensitivity and specificity of blca-1 and BTA in patients with myometrial invasive bladder cancer were compared by using the Fisher exact probability method. The results showed that the sensitivity of ELISA was similar (P0.05), and the specificity of urine blca-1 was significantly higher than that of urine BTA (P0.05). Meanwhile, the sensitivity of urine BLCA-1 to detect low grade muscle invasive bladder cancer was significantly higher than that of urine BTA (P0.05), which was 80% (4/5) and 20% (1/5), respectively, and the specificity difference was not statistically significant (P0.05), which was 88% (15/17) and 94% (16/17), respectively. The accuracy, positive predictive value, negative predictive value and Youden index of urine BLCA-1 for the diagnosis of bladder cancer were higher than that of urinary BTA. Wang Xiaopeng and others showed that the sensitivity of ELISA to detect BLCA-4 in bladder was 91% (22/24) and specificity was 100%, but the expression of BLCA-4 in serum was of no diagnostic value. The sensitivity and specificity of urinary BLCA-4 were analyzed by Fisher accurate probability method.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R737.14

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