PCA3在前列腺癌患者前列腺按摩后尿液中的表达

目的检测PCA3在前列腺癌（PCa）患者前列腺按摩后尿液中的表达情况，并讨论其临床意义。方法从56例PCa患者、23例良性前列腺增生（BPH）患者和9例健康男性志愿者（对照组）前列腺按摩后初始尿液中分离细胞，采用RT—PCR方法检测其中PCA3的表达情况。结果BPH和对照组未见PCA3阳性表达，而PCa患者PCA3阳性率为81．3％（39／48），两者差异具有统计学意义（P〈0．01）。结论前列腺按摩后尿液中PCA3的检测有望成为早期诊断PCa的一种较敏感的方法，也有望成为PCa治疗后监测的一种方法。     

前列腺癌患者前列腺液中DD3 mRNA检测及临床意义

目的检测前列腺癌（PCa）患者前列腺液中DD3基因的表达,讨论其临床意义。方法收集31例PCa患者、59例前列腺增生（BPH）患者前列腺液,离心取细胞沉淀物,提取总RNA,采用逆转录-聚合酶链反应（RT-PCR）琼脂糖电泳法方法检测其中DD3 mRNA的水平。同时用2χ检验分析比较不同Gleason分级的PCa患者DD3 mRNA阳性率是否存在差异。结果 PCa患者前列腺液中DD3 mRNA阳性率（77.42%）明显高于BPH患者（5.08%）,两者具有显著性差异（P〈0.01）。DD3 mRNA表达阳性率诊断PCa的灵敏度为77.42%,特异度为94.91%,不同Gleason分级的PCa患者DD3 mRNA表达阳性率无显著统计学差异（P〉0.05）。结论检测前列腺液中DD3 mRNA诊断PCa是一种有效的无侵入性的良好方法。     

DD3 mRNA在前列腺癌组织中定量表达分析

目的:探讨DD3基因在前列腺组织中表达的临床意义。方法:用荧光定量RT-PCR方法对21例前列腺癌(PCa)组织、27例非前列腺部位的肿瘤组织、39例良性前列腺增生(BPH)组织和15例正常前列腺组织中的DD3的表达进行了定量分析,用ROC曲线对DD3 mRNA诊断PCa的性能进行了分析。结果:27例非前列腺组织中均未检测到DD3基因的表达。DD3在PCa组、BPH组和正常前列腺组表达量的中位数分别为7.2×106、2.5×104、1.5×104拷贝/mg组织。PCa组较BPH组和正常前列腺组DD3的表达量显著增高(P<0.01),而BPH组和正常前列腺组间则差异无显著性(P>0.05)。DD3表达量与临床分期和分化程度之间均无明显相关性。DD3 mRNA曲线下面积(AUC-ROC)为0.937(95%CI:0.879～0.995)。当临界值为1.4×105拷贝/mg组织时,灵敏度、特异度、准确度、阳性预测值(PPV)、阴性预测值(NPV)、阳性拟然比(+LR)、阴性拟然比(-LR)分别为90.5%、85.0%、86.7%、76.0%、94.3%、6.03和0.11。结论:DD3 mRNA的表达仅限于前列腺组织,具有良好的组织特异性。DD3 mRNA表达在PCa组织中显著升高,在正常前列腺和BPH组织中差异无显著性。DD3 mRNA可作为PCa诊断的良好标志物,在PCa早期诊断、微转移诊断、预后判断、指导治疗等方面也具有潜在的应用价值。     

前列腺癌特异性基因DD3在前列腺癌患者外周血中表达的意义

目的探讨前列腺癌特异性基因DD3在前列腺癌患者外周血中表达及临床意义.方法采集44例中晚期前列腺癌（PCa）患者、20例良性前列腺增生（BPH）患者及18例健康男子外周血,分离单个核细胞,提取总RNA.RT-PCR琼脂糖电泳法检测DD3 mRNA,以β-actin为内参照,以出现311-bp和184 bP片段为DD3 mRNA阳性,仅出现184 bp片段为DD3 mRNA阴性. 结果 20例BPH患者和18例健康青年男子外周血标本中均无DD3 mRNA表达,44例PCa血标本中DD3mRNA表达13例（29.5%）.B期、C期及D期患者DD3 mRNA阳性率分别为0%（0/3）、18%（2/11）及37%（11/30）,各期之间差异无统计学意义（P＞0.05）.PCa伴骨转移患者外周血中DD3 mRNA阳性率为44%（10/22）,而无骨转移者其阳性率为14%（3/22）,组间差异有统计学意义（P＜0.05）.PCa未治疗组、去势治疗组的DD3 mRNA阳性率分别为64%（7/11）、18%（6/33）,组间差异有统计学意义（P＜0.01）.结论 PCa患者外周血中可检测到DD3 mRNA表达,DD3 mRNA可望作为诊断PCa血行转移和治疗监测的良好标志物.     

PSA值增高患者前列腺按摩后尿液中DD3mRNA定量检测无侵入性诊断前列腺癌的意义

目的:探讨PSA值增高患者前列腺按摩后尿液沉渣中DD3mRNA检测在前列腺癌诊断中的应用价值.方法:在前列腺穿刺活检前或术前麻醉后收集患者前列腺按摩后患者的尿液,离心取细胞沉淀物,用逆转录-聚合酶链反应(RT-PCR)法和荧光实时定量逆转录-聚合酶链反应(Real time RT-PCR)方法检测DD3mRNA和PSAmRNA含量,以PSAmRNA作为管家基因校正沉渣中的前列腺细胞,以DD3mRNA/PSAmRNA比值表示DD3mRNA含量.SPSS13.0分析相关数据,以穿刺活检或术后病理检查结果为金标准,同时探讨了前列腺按摩后尿液中DD3Mrna含量与前列腺癌病理分级之间的关系.结果:PSA值增高患者中前列腺癌患者前列腺液DD3mRNA表达量明显高于BPH,差异具有统计学意义.不同病理分级之间DD3mRNA的含量差异无统计学意义(P>0.05).结论:PSA值增高的患者行前列腺按摩后尿液沉渣中DD3mRNA含量检测,较前列腺活检的损伤更小,作为前列腺癌的一种非损伤性诊断方法具有良好的应用前景.     

DD3和PSA基因在前列腺癌组织中的定量表达分析

目的探讨DD3 mRNA和PSA mRNA在前列腺组织中表达的临床意义及诊断前列腺癌(PCa)的价值。方法荧光定量RT—PCR法分析21例PCa组织、39例良性前列腺增生(BPH)组织DD3 mRNA和PSA mRNA的表达,ROC曲线对DD3 mRNA、PSA mRNA和DD3 mRNA／PSA mRNA比值诊断PCa的价值进行分析。结果PCa组织中DD3 mRNA、PSA mRNA表达量和DD3 mRNA／PSA mRNA比值均显著高于BPH组织,差异有统计学意义(P<0．01)。不同临床分期和分化程度之间DD3 mRNA、PSA mRNA和DD3 mRNA／PSA mRNA比值差异无统计学意义(P值均>0．05)。ROC曲线分析结果显示,DD3 mRNA、PSA mRNA和DD3 mRNA／PSA mRNA的曲线下面积分别为0．937、0．755和0．839。当DD3 mRNA、PSA mRNA和DD3 mRNA／PSA mRNA临界值分别为1．4×105拷贝／mg组织、3．0×107拷贝／mg组织和5．0×10-3时,敏感性分别为90．5％、81．0％和81．0％,特异性分别为85．0％、62．0％和66．7％。若将DD3 mRNA和PSA mRNA联合用于PCa的诊断,其特异性与DD3 mRNA相同,为85．0％,敏感性可达100．0％。结论PCa组织中DD3 mRNA和PSA mRNA表达量均增加,但组织中DD3 mRNA的定量检测更具诊断价值,联合检测有利于提高诊断敏感性,对PCa的诊断具有一定临床应用价值。     

uPCA3mRNA及PCA3mRNA/D定量检测对PSA灰区前列腺癌诊断的临床研究

目的:研究前列腺按摩后尿液中PCA3mRNA及PCA3mRNA密度的定量检测在前列腺特异性抗原(PSA)灰区前列腺癌(PCa)临床诊断中的应用价值。方法:选择sPSA在4~10ng/ml的BPH或PCa患者200例,收集其前列腺按摩后尿液,离心取细胞沉淀物,提取总RNA,用实时荧光定量RT-PCR方法检测PSAmRNA、PCA3mRNA的含量,以PSAmRNA来校正尿液中的前列腺癌细胞。以PCA3mRNA/PSAmRNA表示PCA3mRNA的含量,经直肠B超测定前列腺体积,计算PSAD和PCA3mRNA密度(PCA3mRNA/D)。以穿刺活检或术后病理检查结果为金标准,用ROC曲线对PCA3mRNA及PCA3mRNA/D诊断PSA灰区PCa的性能进行评价。结果:PCa组的tPSA、PSAD均较BPH组升高(P0.05或0.01)。PCa组PCA3mRNA(P=0.009)含量及PCA3mRNA/D比值(P=0.005)均高于BPH组;以2.01为截断值时,尿PCA3mRNA/D比值诊断PCa的ROC曲线下面积(AUC)为0.903(95%CI:0.790~0.869),其诊断PCa的敏感度和特异度分别为93.3%和66.7%。尿PCA3mRNA/PSAmRNA比值诊断PCa的ROC曲线下面积(AUC)为0.841(95%CI:0.666~0.997),以0.27为截断值时,其诊断PCa的敏感度和特异度为86.5%和77.5%。结论:PCA3mRNA含量和PCA3mRNA/D两个指标定量检测,可以显著提高检出PSA灰区PCa的敏感度,可用于早期PCa的诊断。其中PCA3mRNA/D具有更好的诊断效能。     

环介导等温扩增法检测血浆游离DD3mRNA在前列腺癌诊断中的价值

目的：采用环介导等温扩增技术（loop-mediated isothermal amplification,LAMP）技术进行前列腺癌（PCa）特异基因DD3检测,探讨其诊断PCa的可行性.方法：选择血清PSA异常者60例,行前列腺穿刺活检的同时留取血标本;另留取15例正常男性血进行DD3检测,将DD3检测结果和前列腺病理检查结果进行比较,探讨血DD3mRNA在PCa诊断中的临床应用价值.结果：60例血清PSA异常者中,27例病理检查诊断为PCa,这些患者均检测到DD3表达,33例前列腺增生患者中有1例DD3阳性表达,15例正常对照组均未见DD3阳性表达.结论：LAMP法检测DD3与病理检测具有极高的一致性,方法简单、快速,可大大减少不必要的前列腺穿刺活检,对PCa的诊断具有重要的临床指导价值.     

不同前列腺组织中雄激素受体的表达研究

目的:研究雄激素受体(AR)在正常前列腺、良性前列腺增生(BPH)和前列腺癌(PCa)组织中的表达,探讨AR与BPH和PCa的关系。方法:采用实时定量PCR、免疫荧光和组织蛋白电泳方法,分析15例正常前列腺、20例BPH与40例PCa标本中AR的表达情况。结果:实时定量PCR和组织蛋白电泳检测BPH组织与正常前列腺组织中AR的表达量差异无统计学意义(P>0.05)。但免疫荧光检测发现BPH组织中AR蛋白表达量增高。3种方法检测PCa组织中AR表达量较正常前列腺组织和BPH组织增高(P<0.05)。高分化PCa的AR表达比低分化PCa高(P<0.05)。随着临床分期的增高,AR的表达降低(P<0.05),激素非依赖性前列腺癌(HRPC)组织中AR表达最低。结论:AR在PCa组织中的表达较正常前列腺和BPH组织中增高,AR的表达与PCa的分级、分期相关。     

血管内皮生长因子受体激酶插入嵌合受体在前列腺癌及良性前列腺增生组织中的表达

目的:探讨血管内皮生长因子受体KDR在PCa组织中的表达及其与PCa病理学分级之间的关系。方法:用KDR多克隆抗体对48例PCa组织和20例BPH组织石蜡包埋切片进行免疫组化染色。分析KDR在PCa、BPH组织中的表达情况以及表达强度与PCa组织病理学分级之间的关系。结果:KDR在73%PCa组织、30%BPH组织中呈不同程度的阳性表达。在PCa组织中呈明显高表达(P<0.05),表达强度与前列腺癌的Gleason评分无相关性(r=0.09,P>0.05)。结论:KDR在PCa组织中的表达明显增高,有望成为治疗PCa的一个新靶点。     

ENDOTHELIAL CELL PROLIFERATION ACTIVITY IN BENIGN PROSTATIC HYPERPLASIA AND PROSTATE CANCER: AN IN VITRO MODEL FOR ASSESSMENT

Purpose

Urinary excretion of several pro-angiogenic and antiangiogenic substances has been correlated with malignant tumor growth. The aim of this study was to assay angiogenic activity in urine from patients with cancer of the prostate and benign prostatic hyperplasia (BPH).

Materials and Methods

Urine specimens from 22 healthy male volunteers (control), 33 patients with BPH and 29 with organ confined prostate cancer were analyzed for angiogenic activity in a bovine capillary endothelial cell proliferation assay. In parallel the concentration of basic fibroblast growth factor and vascular endothelial growth factor was determined by enzyme immunoassay in the corresponding urine specimens.

Results

Urine samples from patients with BPH and prostate cancer increased bovine capillary endothelial cell proliferation by 13.1% and 15.1%, respectively, whereas urine from the control group showed a significantly lower angiogenic activity, increasing endothelial cell proliferation by only 0.7% (p = 0.001). Urinary basic fibroblast growth factor and vascular endothelial growth factor were highest in patients with BPH and lowest in the group with prostate cancer (p = 0.0001).

Conclusions

Urine from patients with BPH and prostate cancer stimulates endothelial cell proliferation activity. The degree of endothelial cell stimulation does not correlate with the concentration of basic fibroblast growth factor or vascular endothelial growth factor. Whether the observed pro-angiogenic activity is due to an increased production or release of (an) other angiogenic factor(s) and/or loss of (an) angiogenesis inhibitor(s), deserves further investigation.     

DD3基因不同外显子在前列腺癌组织中的特异性表达

目的 探讨DD3基因不同外显子在前列腺癌组织中的特异性表达。方法 根据基因库所收录的DD3 mRNA序列，设计跨越不同外显子的3对DD3 mRNA序列特异的引物，应用逆转录-聚合酶链反应（RT-PCR）对61例前列腺组织和70例前列腺外的其他肿瘤组织及癌旁组织进行DD3 mRNA检测，并对前列腺癌组织DD3 mRNA进行全序列分析。结果 跨越DD3 mRNA外显子1与3、外显子1与4及外显子3与4之间设计的引物所检测DD3 mRNA结果完全一致，21例前列腺癌组织与1例前列腺上皮内肿瘤组织中DD3 mRNA均为阳性，39例良性前列腺增生组织和70例其他肿瘤组织及癌旁组织中DD3 mRNA均为阴性。21例前列腺癌组织DD3 mRNA序列之间完全一致（GenBank登陆号为AY894120），与国外报道序列（AF103907）之间有99．8％同源。结论 DD3 mRNA仅在前列腺癌中特异性表达，外显子3和外显子4均可作为DD3 mRNA特异的外显子。     

DD3PCA3 RNA analysis in urine--a new perspective for detecting prostate cancer

OBJECTIVES: Serum tPSA lacks specificity. The DD3(PCA3) gene is highly specific for prostate cancer and is detectable in prostate cancer cells shed into urine after rectal palpation. A newly developed nucleic acid sequence based amplification assay (uPM3) for detecting DD3(PCA3) RNA in urine samples was evaluated prospectively in patients referred for prostate cancer detection. METHODS: The uPM3 assay simultaneously detects the relative expression of DD3(PCA3) RNA and PSAmRNA as a marker for prostate cells in urine. Urine samples were collected after attentive digital rectal palpation prior to transrectal guided prostate biopsy. Samples were provided as a single void specimen (20-30 ml), stabilized in phosphate buffer and centrifuged. Lysis was performed on cell pellets DD3(PCA3) RNA and PSAmRNA were extracted and amplification was performed using isothermic nucleic acid based amplification (NASBA). The two targets were detected in real-time using specific beacons as probes in a thermostated spectrofluorimeter. Parameters of the amplification curve were defined after a logistic curve fitting routine and a classification tree model was constructed to predict the outcome of patients (i.e. cancer and non-cancer). RESULTS: 201 patients were included in this prospective study. 158/201 analyzed urine samples contained enough prostate cells sufficient for DD3(PCA3) analysis (79% adequacy rate). Prostate cancer was found in 62 (39%) of the evaluable patients. Overall sensitivity, specificity, positive predictive value and negative predictive value for the uPM3 assay at a cut-off 0.5 probability were 82%, 76%, 67% and 87% respectively as compared to 98%, 5%, 40% and 83% respectively for tPSA (at a cutoff of 2.5 ng/ml). In the tPSA categories <4, 4-10 and >10 ng/ml sensitivity was 73%, 84% and 84% and specificity was 61%, 80% and 70%, respectively. The AUC (area under the curve) was 0.87 (CI 0.81-0.92). CONCLUSION: The uPM3 assay showed excellent clinical performances and a specificity far superior to tPSA.     

前列腺特异膜抗原mRNA的临床应用研究

目的 :通过检测治疗前后的前列腺癌 (PC)病人外周血前列腺特异膜抗原信使核糖核酸 (PSMAmRNA)的表达 ,了解原发性前列腺癌的血行播散情况及为前列腺癌综合治疗提供依据。　方法 :采用巢式逆转录—聚合酶链反应 (RT PCR)检测技术 ,对 31例PC病人外周血PSMAmRNA的表达进行了研究。　结果 :31例样本中 ,17例PC病人 (5 4.8% )PSMAmRNA阳性 ,10例正常人和 2 1例良性前列腺肥大 (BPH)病人全阴性 ;10例伴远处转移的PC病人外周血PSMAmRNA全阳性 ,2 1例检测时尚无转移证据者 33.3% (7/ 2 1)阳性 ;外周血PSMAmRNA阳性率与血清PSA水平密切相关 ;根治术或姑息性治疗前、后外周血PSMAmRNA阳性率差异无显著性。　结论 :外周血PSMAmRNA检测可诊断前列腺癌血行播散及转移 ,可作为预测前列腺癌复发、转移的参考指标 ,对外周血PSMAmRNA阳性病人应在围手术期辅以激素、放疗或免疫等辅助治疗 ,以预防肿瘤复发、转移。     

The association of prostate stem cell antigen (PSCA) mRNA expression and subsequent prostate cancer risk in men with benign prostatic hyperplasia following transurethral resection of the prostate

BACKGROUND: Prior data showed prostate stem cell antigen (PSCA) mRNA expression in benign prostatic hyperplasia (BPH) tissues. The purpose of the present investigation was to determine whether PSCA mRNA expression in resected BPH samples was associated with the subsequent presence of cancer following transurethral resection of the prostate (TURP). METHODS: PSCA in situ hybridization was performed on the TURP-resected tissues from 288 patients, who were histopathologically confirmed BPH without cancer. All these patients were continuously followed for 9-70 months postoperatively. Univariate and multivariate cox regression analyses were used to evaluate the predictive performance of PSCA mRNA for subsequent cancer onset following TURP. RESULTS: PSCA mRNA was detected in 93/288 (32.3%) of the resected BPH specimens, with a mean positive-labeling cells of 23.8%, in which 22 patients (23.7%) were identified as having PCa on follow-up. Of 195 patients with negative expression for PSCA mRNA 2 (1.0%) were subsequently found with PCa. PSCA mRNA expression levels were directly proportional to higher Gleason score and clinical T stage. Univariate and multivariate cox regression analyses demonstrated that only PSCA mRNA expression was predictive of the subsequent cancer development after TURP, however, PSA velocity was an univariately significant but not multivariately significant predictor. CONCLUSIONS: This prospective study identifies PSCA mRNA in BPH as a significant predictor of cancer development after TURP, suggesting that PSCA may be used to identify patients who are at high risk for subsequent cancer onset following TURP for BPH and the PSCA test may be useful when applied for repeat biopsies.