BLCA-4在浸润性膀胱癌患者体液和组织中表达的研究
发布时间：2018-08-26 16:19
【摘要】：目的： 膀胱癌是我国泌尿系统常见的恶性肿瘤之一，无痛性肉眼血尿为最常见的主诉。目前膀胱镜检查和尿脱落细胞检查是诊断和随访膀胱癌的最常用方法，但膀胱镜检查为有创检查，费用较高且给患者（尤其男性）带来极大的不适，还可能造成泌尿系感染。而尿脱落细胞学检查需要在恰当的时间收集尿液才能提高诊断率，虽然特异性较高，但敏感度较低，极易出现假阴性，会造成漏诊。近些年来，随着蛋白组学、基因组学和分子生物学等学科的迅猛发展,许多新的膀胱肿瘤瘤标得以发现,如miRNA、FDP(纤维蛋白原降解产物)、BTA (膀胱肿瘤抗原)、ImmunoCyt（免疫荧光细胞学）、UroVysion、核基质蛋白（NMP22）等。但是，这些瘤标对膀胱癌诊断的敏感性、特异性均不尽人意，有的实验要求的条件极为苛刻、价格昂贵、假阳性率高、容易受到主观因素的影响等，诸多问题使得这些瘤标不能广泛的应用于临床实践。 BLCA-4（bladder cancer specificity nuclear matrix protein4）是膀胱癌特异性核基质蛋白，BLCAS家族中的一种，只存在于膀胱癌组织及癌旁组织中，与癌细胞的增殖、存活和血管的生成等密切相关。研究显示尿中BLCA-4对膀胱癌诊断有着极高的敏感性和特异性，且不会受其他泌尿系疾病（如结石、前列腺增生等良性疾病）影响。但该项研究尚不深入，目前只有以尿液为基础的关于敏感性及特异性的研究，而BLCA-4在血清中的表达水平和浸润性膀胱癌组织中表达的研究尚未见报道。本实验拟用免疫组化、ELISA、Western Blotting方法检测BLCA-4在浸润性膀胱肿瘤患者、泌尿系良性疾病和正常人的体液和组织中的表达及其水平，评估BLCA-4与浸润性膀胱肿瘤生物学活性之间的关系，探讨BLCA-4在膀胱肿瘤诊断中的价值，为BLCA-4作为新的膀胱癌标记物的临床应用提供实验依据。 方法： 用ELISA法、免疫组织化学法和Western Blotting法分别检测BLCA-4在实验组（72例浸润性膀胱癌患者的血液、尿液和膀胱组织）、对照组（包括78例前列腺增生患者的血液、尿液和膀胱组织，44例体检正常者和34例泌尿系结石患者的血液和尿液）中的水平及表达情况。实验结果采用SPSS17.0统计软件进行分析，采用秩转换非参数检验、单因素方差分析以及T检验，P值小于或等于0.05将被认为所检验的差别有统计学意义。 结果： ELISA方法检测结果显示：实验组浸润性膀胱癌患者尿液中BLCA-4含量中位数为1.593，增生组含量为0.319，结石组含量为0.238，正常组为0.194，浸润性膀胱癌患者尿液BLCA-4蛋白含量显著高于其余三组（P0.05），增生组BLCA-4含量较正常对照组升高（P0.05）；BLCA-4在T2a含量中位数为1.809，T2b中含量为1.675，T3+T4中含量为1.982，在中高分化膀胱癌尿液BLCA-4为1.416、低分化膀胱癌尿液BLCA-4为1.817，膀胱癌患者尿液BLCA-4含量在不同分级分期之间差别无统计学意义（P0.05）；肿瘤直径大小＜2cm BLCA-4含量为0.966和肿瘤直径大小≥2cm含量为1.809，膀胱癌患者尿液BLCA-4含量与肿瘤大小有关（P0.05）。确定临界值为0.620ng/ml时，尿BLCA-4检测浸润性膀胱癌的灵敏度、特异度最佳，分别为91.6%（22/24）和100%；实验组浸润性膀胱癌患者血清中BLCA-4含量中位数为5.808，增生组、结石组、正常组患者血清中BLCA-4的含量分别为5.718、5.076、4.995，结果分析无统计学意义（P0.05）。（见Fig1、2、3、4，Tab1、2、3)。 免疫组织化学检测结果显示：浸润性膀胱癌组织中BLCA-4的表达阳性率（77.8%）明显高于对照组前列腺增生患者膀胱组织中的表达阳性率（2.56%），BLCA-4在中高分化和低分化癌组织中表达阳性率分别为65%和93.7%，BLCA-4在T2a、T2b和T3-T4中表达阳性率分别为64.3%、72.7%和100%，BLCA-4的表达与肿瘤分级（P＜0.05）分期（P＜0.05）有相关性；癌旁组织（距切缘2cm内）中BLCA-4表达阳性率为50%（36/72）；与患者年龄（P=0.801）、性别（P=0.289）、肿瘤数量（P=0.526）、肿瘤大小（P=0.076）、手术方式（P=0.848）无明显相关性（P＞0.05）(见Fig7、8、9，Tab5)。 Western-Blotting结果显示：浸润性膀胱癌组织中BLCA-4表达为0.828±0.267，癌旁组织表达为0.591±0.259，前列腺增生患者组织中表达为0.242±0.116，BLCA-4在在浸润性膀胱癌组织中表达较高，在癌旁组织中也有表达，在前列腺增生患者膀胱组织中基本无表达，三者表达差异有统计学意义（P＜0.05）(Fig5、6，Tab4)。 结论： 1. BLCA-4在浸润性膀胱癌患者尿液中的表达有较高的敏感性（91.6%）和特异性（100%），但其在浸润性膀胱癌患者血液中的表达在本实验中未发现差异有统计学意义。 2. BLCA-4在浸润性膀胱癌患者尿液中的表达与肿瘤直径大小存在一定相关性。 3. BLCA-4在浸润性膀胱癌组织中表达的水平与膀胱癌的分级、分期有相关性，与患者年龄、性别、肿瘤数量、肿瘤大小和手术方式无明显相关性。 4.BLCA-4在在浸润性膀胱癌组织中表达较高，在癌旁组织中也有表达，，在前列腺增生患者膀胱组织中基本无表达，三者表达差异有统计学意义。
[Abstract]:Objective:
Bladder cancer is one of the common malignant tumors of the urinary system in China. Painless gross hematuria is the most common complaint. Cystoscopy and urinary exfoliated cells are the most common methods for the diagnosis and follow-up of bladder cancer. However, cystoscopy is invasive, costly and brings great discomfort to patients (especially men), and may also be manufactured. In recent years, with the rapid development of proteomics, genomics and molecular biology, many new bladder tumors have emerged. However, the sensitivity and specificity of these tumor markers to the diagnosis of bladder cancer are not satisfactory, and some of the experimental conditions are very harsh, expensive, high false positive rate, tolerate. These tumor markers can not be widely used in clinical practice because they are susceptible to subjective factors.
BLCA-4 (bladder cancer specific nuclear matrix protein4) is one of the bladder cancer specific nuclear matrix proteins, which only exists in bladder cancer tissues and adjacent tissues, and is closely related to the proliferation, survival and angiogenesis of cancer cells. Studies have shown that urinary BLCA-4 is highly sensitive and specific for the diagnosis of bladder cancer. However, this study is not yet thorough. At present, only urine-based studies on sensitivity and specificity have been conducted. However, the expression of BLCA-4 in serum and in invasive bladder cancer have not been reported. The expression and level of BLCA-4 in the body fluid and tissues of patients with invasive bladder tumor, benign urinary diseases and normal subjects were detected by immunohistochemistry, ELISA and Western Blotting methods. The relationship between BLCA-4 and the biological activity of invasive bladder tumor was evaluated. The value of BLCA-4 in the diagnosis of bladder tumor was discussed. The experimental basis for clinical application of caspin markers is provided.
Method:
BLCA-4 in blood, urine and bladder tissues of 72 patients with invasive bladder cancer, 78 patients with benign prostatic hyperplasia, 44 normal subjects and 34 patients with urinary calculi were detected by ELISA, immunohistochemistry and Western Blotting, respectively. Statistical software SPSS17.0 was used to analyze the experimental results. Rank-switching nonparametric test, one-way ANOVA and T-test were used to analyze the results. The difference between the two tests was considered statistically significant if P value was less than or equal to 0.05.
Result:
The results of ELISA showed that the median content of BLCA-4 in urine was 1.593 in the experimental group, 0.319 in the proliferative group, 0.238 in the calculus group and 0.194 in the normal group. The median content of BLCA-4 was 1.809 in T2a, 1.675 in T2b, 1.982 in T3+T 4, 1.416 in moderately and highly differentiated bladder cancer, 1.817 in poorly differentiated bladder cancer, and no significant difference was found between different stages of bladder cancer (P 0.05). When the critical value was 0.620 ng/ml, the sensitivity and specificity of urinary BLCA-4 for detecting invasive bladder cancer were the best, 91.6% (22/24) and 100% respectively. The serum BLCA-4 levels in patients with invasive bladder cancer in the experimental group were 91.6% (22/24) and 100%, respectively. The number of BLCA-4 in serum was 5.808, 5.718, 5.076, 4.995 in the proliferative group, calculus group and normal group, respectively. The results showed no statistical significance (P 0.05). (See Fig 1, 2, 3, 4, Tab 1, 2, 3).
The positive rate of BLCA-4 expression in invasive bladder cancer (77.8%) was significantly higher than that in control group (2.56%). The positive rate of BLCA-4 expression in moderately well differentiated and poorly differentiated bladder cancer was 65% and 93.7%, respectively. The positive rate of BLCA-4 expression in T2a, T2b and T3-T 4 was positive. The expression of BLCA-4 was correlated with tumor grading (P P > 0.05) (see Fig7,8,9, Tab5).
Western-Blotting results showed that the expression of BLCA-4 in invasive bladder cancer was 0.828.267, 0.591.259 in adjacent tissues, 0.242.116 in benign prostatic hyperplasia, higher in invasive bladder cancer, and higher in adjacent tissues, and basal cells in benign prostatic hyperplasia. There was no significant difference in expression between the three groups (P < 0.05) (Fig5,6, Tab4).
Conclusion:
1. The expression of BLCA-4 in urine of patients with invasive bladder cancer was highly sensitive (91.6%) and specific (100%), but there was no significant difference in the expression of BLCA-4 in the blood of patients with invasive bladder cancer.
2. the expression of BLCA-4 in urine of patients with invasive bladder cancer is related to the diameter of tumor.
3. The expression level of BLCA-4 in invasive bladder cancer was correlated with the grade and stage of bladder cancer, but not with the age, sex, tumor number, tumor size and operation method.
4. The expression of BLCA-4 was higher in invasive bladder cancer tissues and in adjacent tissues, but not in benign prostatic hyperplasia.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14
本文编号：2205464
[Abstract]:Objective:
Bladder cancer is one of the common malignant tumors of the urinary system in China. Painless gross hematuria is the most common complaint. Cystoscopy and urinary exfoliated cells are the most common methods for the diagnosis and follow-up of bladder cancer. However, cystoscopy is invasive, costly and brings great discomfort to patients (especially men), and may also be manufactured. In recent years, with the rapid development of proteomics, genomics and molecular biology, many new bladder tumors have emerged. However, the sensitivity and specificity of these tumor markers to the diagnosis of bladder cancer are not satisfactory, and some of the experimental conditions are very harsh, expensive, high false positive rate, tolerate. These tumor markers can not be widely used in clinical practice because they are susceptible to subjective factors.
BLCA-4 (bladder cancer specific nuclear matrix protein4) is one of the bladder cancer specific nuclear matrix proteins, which only exists in bladder cancer tissues and adjacent tissues, and is closely related to the proliferation, survival and angiogenesis of cancer cells. Studies have shown that urinary BLCA-4 is highly sensitive and specific for the diagnosis of bladder cancer. However, this study is not yet thorough. At present, only urine-based studies on sensitivity and specificity have been conducted. However, the expression of BLCA-4 in serum and in invasive bladder cancer have not been reported. The expression and level of BLCA-4 in the body fluid and tissues of patients with invasive bladder tumor, benign urinary diseases and normal subjects were detected by immunohistochemistry, ELISA and Western Blotting methods. The relationship between BLCA-4 and the biological activity of invasive bladder tumor was evaluated. The value of BLCA-4 in the diagnosis of bladder tumor was discussed. The experimental basis for clinical application of caspin markers is provided.
Method:
BLCA-4 in blood, urine and bladder tissues of 72 patients with invasive bladder cancer, 78 patients with benign prostatic hyperplasia, 44 normal subjects and 34 patients with urinary calculi were detected by ELISA, immunohistochemistry and Western Blotting, respectively. Statistical software SPSS17.0 was used to analyze the experimental results. Rank-switching nonparametric test, one-way ANOVA and T-test were used to analyze the results. The difference between the two tests was considered statistically significant if P value was less than or equal to 0.05.
Result:
The results of ELISA showed that the median content of BLCA-4 in urine was 1.593 in the experimental group, 0.319 in the proliferative group, 0.238 in the calculus group and 0.194 in the normal group. The median content of BLCA-4 was 1.809 in T2a, 1.675 in T2b, 1.982 in T3+T 4, 1.416 in moderately and highly differentiated bladder cancer, 1.817 in poorly differentiated bladder cancer, and no significant difference was found between different stages of bladder cancer (P 0.05). When the critical value was 0.620 ng/ml, the sensitivity and specificity of urinary BLCA-4 for detecting invasive bladder cancer were the best, 91.6% (22/24) and 100% respectively. The serum BLCA-4 levels in patients with invasive bladder cancer in the experimental group were 91.6% (22/24) and 100%, respectively. The number of BLCA-4 in serum was 5.808, 5.718, 5.076, 4.995 in the proliferative group, calculus group and normal group, respectively. The results showed no statistical significance (P 0.05). (See Fig 1, 2, 3, 4, Tab 1, 2, 3).
The positive rate of BLCA-4 expression in invasive bladder cancer (77.8%) was significantly higher than that in control group (2.56%). The positive rate of BLCA-4 expression in moderately well differentiated and poorly differentiated bladder cancer was 65% and 93.7%, respectively. The positive rate of BLCA-4 expression in T2a, T2b and T3-T 4 was positive. The expression of BLCA-4 was correlated with tumor grading (P P > 0.05) (see Fig7,8,9, Tab5).
Western-Blotting results showed that the expression of BLCA-4 in invasive bladder cancer was 0.828.267, 0.591.259 in adjacent tissues, 0.242.116 in benign prostatic hyperplasia, higher in invasive bladder cancer, and higher in adjacent tissues, and basal cells in benign prostatic hyperplasia. There was no significant difference in expression between the three groups (P < 0.05) (Fig5,6, Tab4).
Conclusion:
1. The expression of BLCA-4 in urine of patients with invasive bladder cancer was highly sensitive (91.6%) and specific (100%), but there was no significant difference in the expression of BLCA-4 in the blood of patients with invasive bladder cancer.
2. the expression of BLCA-4 in urine of patients with invasive bladder cancer is related to the diameter of tumor.
3. The expression level of BLCA-4 in invasive bladder cancer was correlated with the grade and stage of bladder cancer, but not with the age, sex, tumor number, tumor size and operation method.
4. The expression of BLCA-4 was higher in invasive bladder cancer tissues and in adjacent tissues, but not in benign prostatic hyperplasia.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14
【参考文献】
相关期刊论文 前1条
1 金美花;德伟林;魏秀鸿;;联合检测尿HAase和NMP-22活性水平在膀胱癌诊断和手术后复发的研究[J];中国实验诊断学;2010年08期
本文编号：2205464
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