BLCA-4在浸潤性膀胱癌患者體液和組織中表達(dá)的研究
發(fā)布時間：2018-08-26 16:19
【摘要】：目的： 膀胱癌是我國泌尿系統(tǒng)常見的惡性腫瘤之一，無痛性肉眼血尿?yàn)樽畛Ｒ姷闹髟V。目前膀胱鏡檢查和尿脫落細(xì)胞檢查是診斷和隨訪膀胱癌的最常用方法，但膀胱鏡檢查為有創(chuàng)檢查，費(fèi)用較高且給患者（尤其男性）帶來極大的不適，還可能造成泌尿系感染。而尿脫落細(xì)胞學(xué)檢查需要在恰當(dāng)?shù)臅r間收集尿液才能提高診斷率，雖然特異性較高，但敏感度較低，極易出現(xiàn)假陰性，會造成漏診。近些年來，隨著蛋白組學(xué)、基因組學(xué)和分子生物學(xué)等學(xué)科的迅猛發(fā)展,許多新的膀胱腫瘤瘤標(biāo)得以發(fā)現(xiàn),如miRNA、FDP(纖維蛋白原降解產(chǎn)物)、BTA (膀胱腫瘤抗原)、ImmunoCyt（免疫熒光細(xì)胞學(xué)）、UroVysion、核基質(zhì)蛋白（NMP22）等。但是，這些瘤標(biāo)對膀胱癌診斷的敏感性、特異性均不盡人意，有的實(shí)驗(yàn)要求的條件極為苛刻、價格昂貴、假陽性率高、容易受到主觀因素的影響等，諸多問題使得這些瘤標(biāo)不能廣泛的應(yīng)用于臨床實(shí)踐。 BLCA-4（bladder cancer specificity nuclear matrix protein4）是膀胱癌特異性核基質(zhì)蛋白，BLCAS家族中的一種，只存在于膀胱癌組織及癌旁組織中，與癌細(xì)胞的增殖、存活和血管的生成等密切相關(guān)。研究顯示尿中BLCA-4對膀胱癌診斷有著極高的敏感性和特異性，且不會受其他泌尿系疾�。ㄈ缃Y(jié)石、前列腺增生等良性疾�。┯绊�。但該項研究尚不深入，目前只有以尿液為基礎(chǔ)的關(guān)于敏感性及特異性的研究，而BLCA-4在血清中的表達(dá)水平和浸潤性膀胱癌組織中表達(dá)的研究尚未見報道。本實(shí)驗(yàn)擬用免疫組化、ELISA、Western Blotting方法檢測BLCA-4在浸潤性膀胱腫瘤患者、泌尿系良性疾病和正常人的體液和組織中的表達(dá)及其水平，評估BLCA-4與浸潤性膀胱腫瘤生物學(xué)活性之間的關(guān)系，探討B(tài)LCA-4在膀胱腫瘤診斷中的價值，為BLCA-4作為新的膀胱癌標(biāo)記物的臨床應(yīng)用提供實(shí)驗(yàn)依據(jù)。 方法： 用ELISA法、免疫組織化學(xué)法和Western Blotting法分別檢測BLCA-4在實(shí)驗(yàn)組（72例浸潤性膀胱癌患者的血液、尿液和膀胱組織）、對照組（包括78例前列腺增生患者的血液、尿液和膀胱組織，44例體檢正常者和34例泌尿系結(jié)石患者的血液和尿液）中的水平及表達(dá)情況。實(shí)驗(yàn)結(jié)果采用SPSS17.0統(tǒng)計軟件進(jìn)行分析，采用秩轉(zhuǎn)換非參數(shù)檢驗(yàn)、單因素方差分析以及T檢驗(yàn)，P值小于或等于0.05將被認(rèn)為所檢驗(yàn)的差別有統(tǒng)計學(xué)意義。 結(jié)果： ELISA方法檢測結(jié)果顯示：實(shí)驗(yàn)組浸潤性膀胱癌患者尿液中BLCA-4含量中位數(shù)為1.593，增生組含量為0.319，結(jié)石組含量為0.238，正常組為0.194，浸潤性膀胱癌患者尿液BLCA-4蛋白含量顯著高于其余三組（P0.05），增生組BLCA-4含量較正常對照組升高（P0.05）；BLCA-4在T2a含量中位數(shù)為1.809，T2b中含量為1.675，T3+T4中含量為1.982，在中高分化膀胱癌尿液BLCA-4為1.416、低分化膀胱癌尿液BLCA-4為1.817，膀胱癌患者尿液BLCA-4含量在不同分級分期之間差別無統(tǒng)計學(xué)意義（P0.05）；腫瘤直徑大�。�2cm BLCA-4含量為0.966和腫瘤直徑大小≥2cm含量為1.809，膀胱癌患者尿液BLCA-4含量與腫瘤大小有關(guān)（P0.05）。確定臨界值為0.620ng/ml時，尿BLCA-4檢測浸潤性膀胱癌的靈敏度、特異度最佳，分別為91.6%（22/24）和100%；實(shí)驗(yàn)組浸潤性膀胱癌患者血清中BLCA-4含量中位數(shù)為5.808，增生組、結(jié)石組、正常組患者血清中BLCA-4的含量分別為5.718、5.076、4.995，結(jié)果分析無統(tǒng)計學(xué)意義（P0.05）。（見Fig1、2、3、4，Tab1、2、3)。 免疫組織化學(xué)檢測結(jié)果顯示：浸潤性膀胱癌組織中BLCA-4的表達(dá)陽性率（77.8%）明顯高于對照組前列腺增生患者膀胱組織中的表達(dá)陽性率（2.56%），BLCA-4在中高分化和低分化癌組織中表達(dá)陽性率分別為65%和93.7%，BLCA-4在T2a、T2b和T3-T4中表達(dá)陽性率分別為64.3%、72.7%和100%，BLCA-4的表達(dá)與腫瘤分級（P＜0.05）分期（P＜0.05）有相關(guān)性；癌旁組織（距切緣2cm內(nèi)）中BLCA-4表達(dá)陽性率為50%（36/72）；與患者年齡（P=0.801）、性別（P=0.289）、腫瘤數(shù)量（P=0.526）、腫瘤大�。≒=0.076）、手術(shù)方式（P=0.848）無明顯相關(guān)性（P＞0.05）(見Fig7、8、9，Tab5)。 Western-Blotting結(jié)果顯示：浸潤性膀胱癌組織中BLCA-4表達(dá)為0.828±0.267，癌旁組織表達(dá)為0.591±0.259，前列腺增生患者組織中表達(dá)為0.242±0.116，BLCA-4在在浸潤性膀胱癌組織中表達(dá)較高，在癌旁組織中也有表達(dá)，在前列腺增生患者膀胱組織中基本無表達(dá)，三者表達(dá)差異有統(tǒng)計學(xué)意義（P＜0.05）(Fig5、6，Tab4)。 結(jié)論： 1. BLCA-4在浸潤性膀胱癌患者尿液中的表達(dá)有較高的敏感性（91.6%）和特異性（100%），但其在浸潤性膀胱癌患者血液中的表達(dá)在本實(shí)驗(yàn)中未發(fā)現(xiàn)差異有統(tǒng)計學(xué)意義。 2. BLCA-4在浸潤性膀胱癌患者尿液中的表達(dá)與腫瘤直徑大小存在一定相關(guān)性。 3. BLCA-4在浸潤性膀胱癌組織中表達(dá)的水平與膀胱癌的分級、分期有相關(guān)性，與患者年齡、性別、腫瘤數(shù)量、腫瘤大小和手術(shù)方式無明顯相關(guān)性。 4.BLCA-4在在浸潤性膀胱癌組織中表達(dá)較高，在癌旁組織中也有表達(dá)，，在前列腺增生患者膀胱組織中基本無表達(dá)，三者表達(dá)差異有統(tǒng)計學(xué)意義。
[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.
【學(xué)位授予單位】：承德醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.14
本文編號：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.
【學(xué)位授予單位】：承德醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.14
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 金美花;德偉林;魏秀鴻;;聯(lián)合檢測尿HAase和NMP-22活性水平在膀胱癌診斷和手術(shù)后復(fù)發(fā)的研究[J];中國實(shí)驗(yàn)診斷學(xué);2010年08期
本文編號：2205464
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