Atg5与P504S在82例前列腺癌中的表达研究

摘 要：［目的］ 探讨Atg5与P504S在前列腺癌中的表达情况。［方法］ 收集前列腺癌（PCa）、良性前列腺增生症（BPH）及前列腺上皮内瘤（PIN）的组织标本共127例,其中PCa组织标本82例,BPH组织标本25例,PIN组织标本20例。采用免疫组化法检测Atg5与P504S在不同病变组织中的表达情况。［结果］ Atg5与P504S在PCa组织中的阳性率均显著高于BPH及PIN组织,差异具有统计学意义（P<0.05）。P504S的阳性表达程度与病理分级、临床分期以及肿瘤转移有关,而与患者年龄无关。Atg5的表达与年龄、病理分级、临床分期以及肿瘤转移均无显著相关性（P>0.05）。Atg5与P504S在PCa组织中的表达呈正相关（P<0.05）。［结论］ Atg5与P504S的异常表达与前列腺癌密切相关,且P504S与前列腺癌进展呈正相关,在前列腺癌诊断中具有较高的临床价值。     

前列腺特异性抗原人类激肽释放酶2和前列腺特异的膜抗原在前列腺癌患者外周血表达的临床意义

目的　探讨检测前列腺癌微转移的灵敏和特异性指标。方法　从 5 1例前列腺癌、33例前列腺增生 (BPH)患者及 32名正常人的外周血中分离单个核细胞 ,用巢式RT PCR方法检测其中前列腺上皮细胞前列腺特异性抗原 (PSA)、人类激肽释放酶 2 (hK2 )和前列腺特异的膜抗原 (PSMA)的表达。结果　PSA、hK2和PSMA在前列腺癌患者外周血中检出的阳性率分别为 5 2 .9%、4 3.1%和6 4 .7% ;正常人和BPH患者假阳性率分别为 6 .2 %、7.7%和 4 .6 % ,3项指标差异均有显著性 (P <0 .0 1)。各临床分期 (局限癌、侵袭性癌和转移癌 )间 ,PSA和hK2的阳性检出率差异无显著性 ;PSMA在各期前列腺癌中阳性检出率均较PSA和hK2高 ,且随临床分期进展 ,其阳性检出率亦增加 (P <0 .0 5 )。结论PSMA对前列腺癌诊断、治疗方案的选择及预后评估较PSA和hK2有更大的价值     

前列腺癌中P504S、34βE12、p63的检测及意义

目的：探讨P504S、p63、34βE12在前列腺腺癌中的表达和诊断意义。方法：应用免疫组化法观察30例前列腺癌（PCa）、49例良性前列腺增生（BPH）、6例不典型腺瘤样增生（AAH）和29例前列腺上皮内瘤变（PIN）组织中P504S、p63、34βE12的表达。结果：28例PCa阳性表达P504S,1例灶性阳性,5例PIN灶性阳性表达P504S;P504S在PCa组阳性明显高于BPH、AAH、和PIN组（P〈0.01）,灶性阳性表达在两组之间无明显差异（P〉0.05）。1例PCa灶性阳性表达p63和34βE12,49例BPH、4例AAH和26例PIN为p63和34βE12阳性,2例AAH和3例PIN为灶性阳性,PCa组p63和34βE12阳性表达明显低于其他病变组（P〈0.01）,灶性阳性表达在两组之间无明显差异（P〉0.05）。结论：P504S是前列腺癌敏感而特异性的标记物,合理利用P504S、p63、34βE12的检测,可提高前列腺癌的诊断准确率和检出率。     

前列腺癌中BAG-1、bcl-2及bax的表达及意义

目的：检测凋亡抑制因子BAG-1在前列腺癌组织（PCa）中的表达，探讨其与前列腺癌发生发展的关系及其与bcl-2和bax表达的关系。方法：采用免疫组织化学染色法检测BAG-1、bcl-2、bax在10例前列腺增生组织（BPH）、45例PCa组织中的表达。结果：BAG-1在BPH、PCa中的阳性表达率分别为20％（2／10）、91．1％（41／45）。PCa中BAG-1表达水平明显高于BPH（P〈0．05），且在癌组织中与肿瘤临床分期（P〈0．01）、病理分级（P〈0．01）呈正相关。BAG-1与bcl-2在PCa中的表达正相关（P〈0．05）。BAG-1与bax在PCa中的表达也呈正相关（P〈0．05）。结论：BAG-1基因可能通过抑制凋亡在PCa的发生发展中发挥重要作用，且其表达与bcl-2、bag的异常表达密切相关     

CCNG2在前列腺癌中的表达及其临床意义

[目的]探讨细胞周期蛋白G2（CCNG2）在前列腺癌中的表达及意义。[方法]采用免疫组织化学方法、Western blot检测85例前列腺癌组织及距其癌组织边缘2cm以上镜下未见癌浸润的正常组织中CCNG2蛋白的表达情况。[结果]免疫组织化学结果表明,CCNG2蛋白在前列腺癌组织及正常前列腺组织中的表达阳性率分别为31.8%、96.5%,差异有统计学意义（P〈0.05）。Western blot结果表明,CCNG2蛋白在前列腺癌组织的相对表达量较正常前列腺组织的相对表达量明显降低,差异具有统计学意义（P〈0.05）。CCNG2蛋白表达与前列腺癌患者年龄、肿瘤大小、PSA水平无关（P〉0.05）;而与Gleason score、淋巴结转移以及肿瘤临床分期有关（P〈0.05）。[结论]前列腺癌组织中CCNG2蛋白表达明显降低,且与前列腺癌Gleason score、淋巴结转移、临床分期有关。     

前列腺癌组织p504S和p63与血清PSA表达水平相关性分析

目的:探讨p504S、p63在前列腺癌(PCa)组织中的表达与血清前列腺特异性抗原(PSA)的关系。方法:采用免疫组织化学方法检测p504S、p63在前列腺增生(BPH)、前列腺上皮内瘤变(PIN)和前列腺癌(PCa)组织中的表达,并探讨前列腺癌(PCa)组织中p504S表达与血清PSA水平的相关性。结果:18例PCa组织中,p504S阳性表达16例(88.89%);28例PIN中,p504S阳性表达6例(21.43%);124例BPH中,p504S弱阳性表达8例(6.45%),p504S在PCa组织中的阳性表达率明显高于PIN与BPH组织,P<0.05。18例PCa组织中,p63呈不连续阳性表达2例(11.11%);28例PIN中,p63阳性表达24例(85.71%);124例BPH中,p63阳性表达116例(93.55%),p63在PCa组织中的表达阳性率明显低于BPH与PIN组织,P<0.05。p504S在血清PSA水平<4 ng/mL的8例PCa中7例阳性表达,在>4 ng/mL的10例PCa中9例阳性表达,两者比较差异无统计学意义,P>0.05。结论:对血清PSA>4 ng/mL、临床疑为PCa的病例,可行穿刺活检联合免疫组织化学方法明确诊断。     

AMACER CK34βE12在前列腺穿刺活检组织中的表达

目的：探讨AMACER和CK3413E12在前列腺穿刺活检组织中的表达，评估其在前列腺癌诊断上的意义．方法：应用免疫组织化学法检测178例前列腺穿刺活检组织中AMACER和CK3413E12表达情况，其中前列腺腺癌（Cap）69倒、良性前列腺增生（BPH）101例、前列腺上皮内组织瘤（PIN）5例、不典型小腺体增生（ASAP）3例．结合临床资料进行分析、，结果：AMACER在微粒穿刺活检组织中CaP阳性表达率为100％，其中弥漫阳性率（≥＋＋＋）为98．6％；BPH中（-）84例（83．2％），（＋）17例（15．8％），（＋＋）1例（1．0％）；PIN和ASAP均为阴性表达。CK3413E12在全部CaP和ASAP病例中为阴性表达，在BPH和PIN病例中均为阳性表达．结论：AMACER是前列腺腺癌的特异性肿瘤标记物，将其与CK3413E12联合应用于穿刺活检所获得前列腺微粒组织的检测，可以确诊前列腺癌，提高早期癌和小体积癌（〈0．5ml）的检出率。     

前列腺干细胞抗原在前列腺组织的表达及意义

目的演探讨前列腺干细胞抗原(prostatestemcellantigen,PSCA)在前列腺组织中的表达及其意义。眼方法演应用核酸分子原位杂交穴ISH雪技术,对PSCAmRNA在26例前列腺癌、20例良性前列腺增生(BPH)和9例正常前列腺(NP)标本中的表达水平进行检测及定位。眼结果演PSCAmRNA在NP、BPH及前列腺癌组织表达阳性率分别为66.7%、70.0%及84.6%,强阳性率分别为0、10.0%及57.7%。NP与BPH组织表达水平无显著差异(P>0.05),其表达定位于前列腺上皮的基底细胞层;PSCAmRNA在前列腺癌组织主要表达于癌细胞,细胞间质和肌肉组织均无表达。前列腺癌与NP、BPH组织表达水平均有显著差异(P<0.01)。PSCAmRNA表达水平与前列腺癌临床分期、病理分级均无相关性(P>0.05)。眼结论演PSCA高表达是前列腺癌的共同特征;PSCA在前列腺癌早期诊断与免疫靶向治疗方面有良好的开发应用前景。     

前列腺癌组织中肿瘤血管生成的研究

为探讨前列腺癌（PCa）细胞转移的机制，应用免疫组织化学SP法在36例良性前列腺增生（BPH）及34例PCa组织中对第Ⅷ因子相关抗原进行表达，计数微血管（MV），发现PCa组织中MV数显著高于BPH组（P＜0.01），浸润性PCa组MV数明显高于局限性PCa组（P＜0．05），伴淋巴结转移者MV数显著高于无淋巴结转移者（P＜0．05），未分化PCa组MV数显著高于高分化组及低分化组（P＜0．05），提示肿瘤血管生成与PCa分期、分级密切相关，癌细胞的浸润及转移有赖于肿瘤血管的生成。     

前列腺癌中 P504S、34βE12、p63 的检测及意义

目的:探讨P504S、p63、34βE12在前列腺腺癌中的表达和诊断意义.方法:应用免疫组化法观察30例前列腺癌(PCa)、49例良性前列腺增生(BPH)、6例不典型腺瘤样增生(AAH)和29例前列腺上皮内瘤变(PIN)组织中P504S、p63、34βE12的表达.结果:28例PCa阳性表达P504S,1例灶性阳性,5例PIN灶性阳性表达P504S;P504S在PCa组阳性明显高于BPH、AAH、和PIN组(P＜0.01),灶性阳性表达在两组之间无明显差异(P＞0.05).1例PCa灶性阳性表达p63和34βE12,49例BPH、4例AAH和26例PIN为p63和34βE12阳性,2例AAH和3例PIN为灶性阳性,PCa组p63和34βE12阳性表达明显低于其他病变组(P＜0.01),灶性阳性表达在两组之间无明显差异(P＞0.05).结论:P504S是前列腺癌敏感而特异性的标记物,合理利用P504S、p63、34βE12的检测,可提高前列腺癌的诊断准确率和检出率.     

Diagnostic Efficacy of PSMA and PSCA mRNAs Combined to PSA in Prostate Cancer Patients

Background: Serum Prostate-specific antigen (PSA) has been used for screening and diagnosis of prostate cancer (PCa) but it is burdened by its low accuracy, creating a need for reliable diagnostic markers. Despite prostate-specific membrane antigen (PSMA) and prostate stem cell antigen (PSCA) being widely expressed in the tissue of PCa, no definite conclusion regarding their use as clinical biomarkers due to their lacking organ specificity. Therefore, this study aimed to evaluate the peripheral blood levels of PSMA and PSCA mRNAs and examine their diagnostic significance as non-invasive integrated markers.Materials and Methods: 125 subjects were enrolled in this study. They were divided into 25 healthy controls, 25 BPH patients, and 75 PCa patients. The expression levels of PSMA and PSCA were determined using quantitative RT- PCR, in addition to measuring serum PSA.Results: Levels of PSMA and PSCA were over-expressed in PCa patients compared to controls and BPH patients and were found to be associated with increased susceptibility to PCa. Moreover, the diagnostic values of PSMA and PSCA to distinguish PCa patients from BPH patients and controls were inferior to that of PSA. However, the combination of PSMA and PSCA with PSA enhanced the efficacy of the latter.Conclusion: This study suggests that these genes were associated with malignant susceptibility. Concerning the duality of PSMA-PSA or PSCA-PSA, this implies the significance of their investigation together in peripheral blood of prostate patients.     

Use of multiple biomarkers for a molecular diagnosis of prostate cancer

The identification of biomarkers capable of providing a reliable molecular diagnostic test for prostate cancer (PCa) is highly desirable clinically. We describe here 4 biomarkers, UDP-N-Acetyl-alpha-D-galactosamine transferase (GalNAc-T3; not previously associated with PCa), PSMA, Hepsin and DD3/PCA3, which, in combination, distinguish prostate cancer from benign prostate hyperplasia (BPH). GalNAc-T3 was identified as overexpressed in PCa tissues by microarray analysis, confirmed by quantitative real-time PCR and shown immunohistochemically to be localised to prostate epithelial cells with higher expression in malignant cells. Real-time quantitative PCR analysis across 21 PCa and 34 BPH tissues showed 4.6-fold overexpression of GalNAc-T3 (p = 0.005). The noncoding mRNA (DD3/PCA3) was overexpressed 140-fold (p = 0.007) in the cancer samples compared to BPH tissues. Hepsin was overexpressed 21-fold (p = 0.049, whereas the overexpression for PSMA was 66-fold (p = 0.047). When the gene expression data for these 4 biomarkers was combined in a logistic regression model, a predictive index was obtained that distinguished 100% of the PCa samples from all of the BPH samples. Therefore, combining these genes in a real-time PCR assay represents a powerful new approach to diagnosing PCa by molecular profiling. (Supplemental material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html.)     

Correlation of primary tumor prostate-specific membrane antigen expression with disease recurrence in prostate cancer.

PURPOSE: The restricted expression of the surface glycoprotein prostrate-specific membrane antigen (PSMA) to normal prostate tissue, primary and metastatic prostate cancer (PCa), and the neovasculature of various nonprostatic epithelial malignancies has enabled targeting strategies for PCa treatment using anti-PSMA antibodies. EXPERIMENTAL DESIGN: Using prostatectomy specimens, immunohistochemical staining for PSMA (7E11 antibody) was performed on formalin-fixed paraffin-embedded sections of 136 cases of PCa. Cytoplasmic immunoreactivity was scored for intensity and distribution, and results were correlated with tumor grade, pathological stage, DNA ploidy status (Feulgen spectroscopy), and disease recurrence. PSMA mRNA expression in selected primary tumors and metastatic lesions was also detected using in situ hybridization and autoradiography. RESULTS: Generally, PCa cells expressed relatively increased levels of PSMA as compared with benign elements. Among the PCa cases, increased (high) PSMA expression correlated with tumor grade (P = 0.030), pathological stage (P = 0.029), aneuploidy (P = 0.010), and biochemical recurrence (P = 0.001). The mean serum prostate-specific antigen level of 18.28 ng/ml at the time of diagnosis for the PSMA-overexpressing tumors was significantly greater than the mean serum prostate-specific antigen of 9.10 ng/ml for the non-PMSA-overexpressing group (P = 0.006). On multivariate analysis, pathological stage (P = 0.018) and PSMA expression (P = 0.002) were independent predictors of biochemical recurrence. PSMA protein overexpression in high-grade primary PCa tumors and metastatic lesions also correlated with increased PSMA mRNA expression levels using in situ hybridization and autoradiography. CONCLUSIONS: This study demonstrates for the first time that overexpression of PSMA in primary PCa correlates with other adverse traditional prognostic factors and independently predicts disease outcome.     

Expression of prostate specific membrane antigen and three alternatively spliced variants of PSMA in prostate cancer patients

Prostate specific membrane antigen (PSMA) is a folate gamma glutamyl carboxypeptidase that is oriented on the plasma membrane of normal and prostate cancer cells. A cytosolic version of PSMA, PSM', results from alternative splicing of the PSMA gene. Two additional alternatively spliced variants of PSMA, PSM-C and PSM-D, have been described recently. The ratio of PSMA to PSM' mRNA was higher in a small number of prostate cancer specimens compared to normal prostate cancer and benign prostatic hypertrophy (Su et al. Cancer Res 1995;55:1441). The intent of our study was to measure the gene expression of PSMA and the 3 PSMA splice variants in a large number of patient's tissues. A real-time, quantitative PCR assay was developed to quantify PSMA, PSM', PSM-C and PSM-D. Discrimination among the variants was achieved by designing unique primers and TaqMan probes for each gene. Amplification and detection was specific for the desired splice variant and was sensitive to one gene copy per reaction. The assay was used to quantify the gene expression in specimens of normal, benign, primary and metastatic prostate cancer from 72 patients. The mean PSMA expression (relative to 18S rRNA) was 2- to 3-fold lower in normal prostate (n = 4) compared to primary (n = 55, p = 0.31) and metastatic (n = 20, p = 0.33) prostate cancer. There was no difference in the PSMA expression between benign and cancerous prostate tissue from the same patients (n = 35). The ratio of PSMA to PSM' was lowest in the normal prostate and increased with increasing Gleason score (p < 0.001). The increased ratio in these tissues was a reflection of both increasing PSMA levels and decreasing PSM' mRNA. The expression of PSM-C did not differ in any of the tissue categories studied. The expression of PSM-D was similar in normal and primary prostate cancer but was 2-fold higher in lymph node (p < 0.005) and bone metastases (p < 0.05) compared to the primary tumors. Our results of the first detailed quantitative analysis of PSMA mRNA expression in patient's tissues demonstrate that PSMA and the 3 PSMA splice variants are expressed in normal, benign, cancerous and metastatic prostate cancer. We note increased PSMA expression in some malignant tissues, however, these increases are modest in magnitude. We also report that the expression of a novel splice variant, PSM-D, is elevated in prostate cancer metastases.     

68Ga-PSMA PET/CT在前列腺癌复发中的应用进展

 前列腺癌（prostate cancer, PCa）发病率高,根治性治疗后,复发病灶的早期诊断对患者的预后至关重要。前列腺特异性膜抗原（prostate specific membrane antigen,PSMA）是一种在PCa中高表达的跨膜糖蛋白,利用核素⁶⁸Ga标记其小分子抑制剂作为显像剂,进行PET/CT显像,可以得到反映PCa病灶在体内分布的影像。欧美很多国家已经将68Ga-PSMA PET/CT应用于寻找PCa患者复发病灶,并取得了很好的效果,该技术在我国还处于起步阶段。本文综述了多个有关⁶⁸Ga-PSMA PET/CT在前列腺癌复发中的研究进展,旨在提高我们对该项新技术的认识水平。     

长链非编码RNA FAL1在前列腺癌组织中的表达及与预后的关系

目的:检测长链非编码RNA FAL1（long non-coding RNA FAL1，lncRNA FAL1）在前列腺癌(PCa)患者癌组织（Tum）和相邻非癌变组织（Non-tum）中的表达差异及其与预后的关系。方法:回顾性选取我院2015年4月至2017年5月治疗的89例PCa患者作为研究对象，记录患者初诊时的年龄、病理Gleason评分、血清中前列腺特异性抗原（prostate specific antigen，PSA）以及TNM分期等。qRT-PCR检测癌组织或相邻非癌变组织中lncRNA FAL1相对表达水平，根据lncRNA FAL1表达水平的中位值将PCa癌组织组分为低、高表达组，通过Log-Rank检验和Kaplan-Meier生存曲线分析lncRNA FAL1对PCa患者预后的影响。最后，Cox多因素分析评估lncRNA FAL1表达与 PCa 患者临床病理特征和预后的关系。结果:与Non-tum组织相比，Tum组织中lncRNA FAL1的表达水平显著性升高，P<0.01；另外，lncRNA FAL1在组织中的表达水平与PCa患者Gleason评分分级、T分期、淋巴转移及远处转移显著相关，P<0.05；和年龄及PSA等指标无关。lncRNA FAL1高表达组的总体生存时间和无进展生存期均显著降低（P<0.05）；lncRNA FAL1高表达是PCa患者预后的危险因素（RR=1.961，95%CI:0.635~2.107，P=0.012）。结论:lncRNA FAL1在PCa患者癌组织中高表达，是预测PCa预后的危险因素，有望成为PCa治疗新的诊断生物标志物和治疗靶点。     

Detection and characterization of the prostate-specific membrane antigen (PSMA) in tissue extracts and body fluids

The prostate-specific membrane antigen (PSMA) glycoprotein is recognized by the monoclonal antibody (MAb) 7EII -C5.3 as a predominant 100 kDa and minor 180 kDa component in LNCaP cell line extracts and its expression has been shown by immunohistochemistry to be highly restricted to prostate epithelium. The aim of the present study was to utilize Western blot analysis to determine if PSMA could be detected in human tissue extracts and body fluids and if so, which molecular forms were present. PSMA was detected as 120 and 200 kDa bands in normal, benign and malignant prostate tissues and seminal plasma. Further analysis demonstrated that the larger molecular form of PSMA may be a dimer of the lower m.w. species. The PSMA glycoprotein was not detected in the majority of non- prostate tissue extracts examined except for a low yet significant amount in normal salivary gland, brain and small intestine, suggesting that PSMA may not be as prostate-specific as originally thought. Since the prostate-specific antigen (PSA) has been shown to be maximally shed into the serum in high-grade and metastatic prostate carcinomas, it was surprising that PSMA could not be detected in serum by Western blot analysis even in patients with actively progressive metastatic disease. Second generation antibodies generated against different epitopes may be required to determine if PSMA is shed into serum. Our results support the hypothesis that PSMA is a novel prostate biomarker. © 1995 Wiley-Liss, Inc.