尿沉淀细胞中TMPRSS2-ETS融合基因对诊断前列腺癌的意义

目的：检测尿沉淀细胞中TMPRSS2-ETS融合基因对诊断前列腺癌的意义。方法：将2010～2011年我院收治的经病理检查证实为前列腺癌者归为前列腺癌组,经病理检查证实为BPH者归为BPH组。收集两组患者前列腺按摩后首次尿液标本,用FISH检测其TMPRSS2-ERG、TMPRSS2ETVl和TMPRSS2-ETV4融合基因的表达。结果：纳入本次研究的前列腺癌组和BPH组分别为51例和20例,TMPRSS2-ERG（＋）的病例分别为26例（50．98％）和4例（20％）,差异有显著统计学意义（P〈O．05）。其敏感度为50．98％,特异度为80％,假阳性率为20％,假阴性率为49．02％,阳性似然比为2．549,阴性似然比为0．613,Youden指数为0．3098。两组中均未发现有TMPRSS2-ETVl或TMPRSS2-ETV4融合基因阳性患者。结论：用FISH方法检测尿沉淀细胞TMPRSS2-ERG融合基因有助于区分前列腺癌与BPH。TMPRSS2-ETV1和TMPRSS2-ETV4融合基因在前列腺癌中出现率较低,故不推荐用于前列腺癌的诊断检测。     

荧光原位杂交技术在前列腺癌诊断中的临床应用

目的建立TMPRSS2/ETS(ERG、ETV1、ETV4)融合基因的荧光原位杂交(FISH)检测和评估方法,并探讨其对于前列腺癌的诊断价值。方法对50例前列腺癌样本、15例正常前列腺组织及20例良性前列腺增生样本应用三组国产FISH探针顺序检测TMPRSS2/ERG、TMPRSS2/ETV1及TMPRSS2/ETV4融合基因,建立FISH技术诊断前列腺癌的阈值,并计算敏感性、特异性、阳性预测值(+PV)和阴性预测值(-PV)。结果三组探针的联合敏感性达到90%,其中TMPRSS2-ERG检测敏感性为78%,总体特异性为100%,阳性预测值为100%,阴性预测值为87.5%。结论应用FISH技术检测TMPRSS2/ETS(ERG、ETV1、ETV4)融合基因诊断前列腺癌具有较高的敏感性和特异性,显示出良好的临床应用前景。     

前列腺癌融合基因TMPRSS2：ERG的检测及其意义

目的：检测国人前列腺癌患者融合基因TMPRSS2：ERG的发生率以及该融合基因和ERG表达的关系。方法：连续选取经直肠B超前列腺癌穿刺证实的前列腺癌新鲜标本62例、6例高级别前列腺上皮内瘤（HGPIN）标本和9例其他器官恶性肿瘤细胞株,另选取10例BPH组织作为对照,采用RT—PCR检测融合基因,采用real-timeRT-PCR检测ERG表达。结果：本组国人前列腺癌患者融合基因TMPRSS2：ERG的发生率为45．16％,融合基因TMPRSS2：ERG诊断前列腺癌的特异度为93．75％,灵敏度为45．16％。前列腺癌融合基因组ERG基因表达显著高于前列腺癌非融合基因组（t=3．549,P=0．001）。结论：融合基因TMPRsS2：ERG在本组国人前列腺癌患者中的发生率以及在HGPIN病灶的发生率与国外研究报道相似。该融合基因诊断前列腺癌的特异性很高,但灵铡度偏低。融合基因和前列腺癌组织中ERG高表达有关。     

ERG基因及其与前列腺癌关系

因属于ETS转录因子家族,在很多肿瘤中高表达,与肿瘤的血管生成、转移、浸润、抑制凋亡有关,ERG基因在前列腺癌中主要受TMPRSS2-ERG融合基因的调节。近一半以上的前列腺癌病例表达TMPRSS2-ERG融合基因,其中ERG基因可能是前列腺癌的重要标志物,它的检测有助于前列腺癌的诊断和治疗方法的选择。TMPRSS2-ERG融合基因的存在还会影响前列腺癌病人的预后。本文就ERG基因的生物学特征、作用机制、TMPRSS2-ERG的融合机制以及ERG基因与前列腺癌的诊断、治疗和预后的关系作一综述。     

融合基因TMPRSS2:ERG与前列腺癌病理分级关系的研究

目的:研究融合基因TMPRSS2:ERG和前列腺癌病理分级的关系。方法:选取前列腺癌的穿刺标本62例为病例组,同时选择10例良性前列腺增生(BPH)患者为对照组,同时纳入9株前列腺癌细胞株作为对照,采用巢式RT-PCR检测融合基因TMPRSS2:ERG,比较融合基因阳性和阴性患者Gleason评分的差异,Logistic回归法分析TMPRSS2:ERG和前列腺癌病理特征的关系。结果:62例前列腺癌患者中有28例检测出TMPRSS2:ERG融合基因,阳性率为45.16%;10例BPH和9株癌细胞株中均未检测出该融合基因。融合基因TMPRSS2:ERG阳性和阴性患者Gleason评分无显著差异(Z=-0.609,P=0.542),但融合基因阳性患者Gleason主评分显著高于阴性患者(Z=-2.600,P=0.009)。单因素Logistic回归分析显示,筛状结构、泡沫状腺体和印戒癌细胞分别与融合基因TMPRSS2:ERG有关联(OR=6.25,P=0.002;OR=6.666,P=0.023;OR=3.24,P=0.035);多因素Logistic回归分析显示,该融合基因和筛状结构有关(OR=3.750,P=0.033)。结论:TMPRSS2:ERG融合基因和前列腺癌中到高级的病理分级有关。     

DOPEY2在TMPRSS2-ERG基因融合阴性前列腺癌中的表达及临床意义

目的探讨DOPEY2在TMPRSS2-ERG基因融合阴性前列腺癌中的表达及其临床意义。方法本研究通过RT-PCR和Western Blot技术检测前列腺癌细胞株VCa P和PC-3中ERG和DOPEY2的表达情况,同时,采用免疫组化技术检测46例前列腺癌根治术后的石蜡标本中ERG和DOPEY2的表达情况,分析两者的表达以及与临床病理特征之间的关系。结果 DOPEY2在TMPRSS2-ERG基因融合阳性前列腺癌细胞株VCa P中的m RNA和蛋白表达水平,均比TMPRSS2-ERG基因融合阴性前列腺癌细胞株PC-3低,且差异均有统计学意义(P0.05)。免疫组化结果显示,DOPEY2在TMPRSS2-ERG基因融合阴性组织中的阳性表达率高于基因融合阳性组(76.5%和33.3%,P0.05),且DOPEY2阳性的患者具有较高的Gleason分级和T分期(P0.05)。ERG蛋白的阳性率为26.1%(12/46),且与低Gleason评分组相关,而与患者的年龄、血浆PSA值,病理T分期和远处转移等参数均无相关性。结论 DOPEY2在TMPRSS2-ERG基因融合阴性前列腺癌中的表达高于TMPRSS2-ERG基因融合阳性前列腺癌,DOPEY2可能作为一个有潜力的分子标记物指导前列腺癌分期及判断预后。     

前列腺癌患者PCA3 mRNA的表达及其临床意义

目的：评价PCA3在前列腺癌诊断、临床分期及Gleason评分中的临床应用价值。方法：选取2005年11月～2006年9月在我院住院的前列腺癌患者56例，其中T2期12例，T3期21例，T4（N0～3，M0～1）期23例；Gleason评分5～7分27例，8～10分29例；BPH患者23例，健康男性9例。分别获取其外周血及前列腺按摩液，采用RT—PCR方法检测两种体液标本中PCA3的表达阳性情况，使用SPSS12．0统计软件包对其结果进行分析。结果：BPH和对照组两种体液标本未见PCA3阳性表达，而PCa患者外周血标本PCA3阳性率为88．9％（48／54），前列腺按摩液标本PCA3阳性率为81．3%（39／48），差异均有统计学意义（P〈0．01）。结论：两种体液标本PCA3的阳性表达有明显的前列腺癌特异性，并且随着前列腺癌的临床分期增高，其阳性率越高，有望成为诊断前列腺癌的新肿瘤标志物和判断预后的指标。     

PCA3 mRNA与PSA mRNA的联合检测对前列腺癌早期诊断的临床意义

目的评价外周血前列腺癌抗原基因-3（prostate cancer antigen3,PCA3 mRNA）和前列腺特异性抗原基因（prostate specific antigen,PSA mRNA）联合检测对前列腺癌（PCa）早期诊断的价值,及决定是否进行前列腺穿刺活检。方法纳入41例PCa和69例前列腺增生（BPH）患者的外周血,采用RT—PCR方法对其PCA3mRNA和PSAmRNA进行检测,通过受试者工作特征（ROC）曲线评价其在预测前列腺穿刺活检结果和PCa早期诊断的价值。结果PCa组外周血PCA3mRNA含量明显高于BPH组[2362（〈30～7421）拷贝／ml比〈35拷贝／ml,z=-6．66,P〈0．01],而PSAmRNA含量也明显高于BPH组[3425（908～36639）拷贝／ml比〈220拷贝／ml,z=-6．40,P〈0．01];ROC曲线显示当PCA3mRNA和PSAmRNA临界值分别为846拷贝／ml和280拷贝／ml时,诊断早期PCa结果为73．2％（30／41）、85．4％（35／41）,特异度分别为87．0％（60／69）、78．3％（54／69）;而联合检测时其敏感度可增至90．2％（37／41）,特异度上升为89．9％（62／69）。结论外周血PCA3mRNA和PSAmRNA检测都是PCa诊断的良好指标,而它们的联合检测可弥补PCA3mRNA敏感性低和PSAmRNA特异性低的缺点,更有利于早期PCa诊断。     

前列腺癌中PIM-1的表达及其临床意义

目的 探讨PIM-1在前列腺癌中的表达及临床意义。方法 逆转录-聚合酶链反应（RT—PCR）半定量分析2例良性前列腺增生（BPH）和5例前列腺癌（PCa）组织标本中PIM-1mRNA表达，免疫组织化学法检测20例BPH、20例高分级前列腺上皮内瘤（HGPIN）和42例PCa组织标本中PIM-1蛋白表达水平，染色结果分为阴性、弱阳性、阳性和强阳性。结果 5例PCa组织PIM-1mRNA表达相对值分别为0．63、0．55、0．42、0．91、0．76，2例BPH中其相对值为0．26、0．27。BPH、HGPIN和PCa组织中PIM-1蛋白阴性表达率分别为60％（12／20）、20％（4／20）和2％（1／42），弱阳性表达率分别为40％（8／12）、20％（4／20）和12％（5／42），阳性列强阳性表达率分别为0（0／20）、60％（12／20）和86％（36／42），PCa中PIM-1蛋白表达水平高于HGPIN和BPH（P值均〈0．05）。PIM-1蛋白表达水平随PCa的临床分期和病理分级增高而增强，在有和没有淋巴结转移PCa组织中PIM-1强阳性表达率分别为70％（7／10）、25％（8／32），差异有统计学意义（P〈0．05）。结论PIM-1高表达可能与PCa发生和发展相关，PIM-1表达水平与PCa分期、Gleason评分呈正相关，可能成为PCa预后判断的肿瘤标志物。     

Molecular profiling of ETS gene rearrangements in patients with prostate cancer registered in REDEEM clinical trial

ObjectiveAndrogen-induced E26 transformation-specific (ETS) gene fusion–positive tumors have been associated with aggressive prostate cancer. The aim is to evaluate the ETS gene rearrangement status on initial biopsy of patients registered in the Reduction by Dutasteride of Clinical Progression Events in Expectant Management trial study and determine if gene fusion status was associated with disease progression.Materials and methodsInitial biopsy material from 146 men registered in Reduction by Dutasteride of Clinical Progression Events in Expectant Management trial study treated with dutasteride (73/146, 50%) and as placebo (73/146, 50%) were reviewed, and ERG and SPINK1 immunohistochemistry was performed. ERG- and SPINK1-negative cancer samples were evaluated for ETV1, ETV4, and ETV5 rearrangements by fluorescence in situ hybridization. Frequency of ETS gene aberrations in both groups was correlated with cancer progression including prostate-specific antigen progression, Gleason progression, and progression-free survival by logistic analysis, pairwise differences, and chunk likelihood ratio tests for the genotype groups.Results and conclusionsOf the 146 patients, 99 (67.8%) (placebo, 51; dutasteride, 48) samples displayed the following Gleason patterns: 3+3 = 6 in 80 (54.8%) (placebo, 39; dutasteride, 41), 3+4 = 7 in 18 (12.3%) (placebo, 11; dutasteride, 7), and 4+4 = 8 in 1(0.68%) (placebo, 1). The remaining 47 samples showed atypical glands in 5 (3.4%) (placebo, 2; dutasteride, 3), HGPIN in 9 (6.1%) (placebo, 5; dutasteride, 4), and benign in 33 (22.6%) (placebo, 15; dutasteride, 18). Immunohistochemistry findings were positive for ERG and SPINK1 in 56 (56%) (placebo, 31; dutasteride, 25) and 9 (6.1%) (placebo, 5; dutasteride, 4) cases, respectively. ETV1 and ETV4 rearrangements were noted in 2 cases (1.4%) (placebo, 1; dutasteride, 1) and 1 (0.7%) (placebo, 1) case, respectively. No significant differences in the incidence of prostate cancer molecular aberrations between the groups were observed. There was no evidence that ETS fusion status was associated with disease progression.     

The utility of ERG/P63 double immunohistochemical staining in the diagnosis of limited cancer in prostate needle biopsies

Diagnosis of limited cancer can be challenging in prostate needle biopsies, and immunohistochemistry is commonly used in such settings. Recently, TMPRSS2:ERG gene rearrangement was found to be highly specific for and detected in approximately 50% of prostate cancer. Positive immunohistochemical staining with a novel anti-ERG antibody highly correlated with TMPRSS2:ERG gene rearrangement status. We developed a double immunohistochemical staining containing both erythroblastosis virus E26 oncogen (ERG) and basal cell marker P63 antibodies and evaluated its use in the diagnosis of limited cancer in prostate needle biopsies. A total of 77 prostate needle biopsies containing cancer occupying <1 mm of the length of only 1 core of the entire biopsy set were stained with the double stain containing ERG and P63 antibodies. ERG positivity and its staining intensity in cancerous and other noncancerous lesions were evaluated. ERG expression was detected in 42% (32 of 77) of cases, with strong, moderate, and weak staining intensity in 72%, 16%, and 12% of cases. The staining was uniform in 84% of cases and heterogeneous in 16% of cases with different staining intensities in >10% of cancerous cells. High-grade prostatic intraepithelial neoplasia was present in 17 cases, and in 5 (29%) cases ERG was positive in high-grade prostatic intraepithelial neoplasia glands, which were all immediately adjacent to or intermingled with ERG-positive cancerous glands. In 4 additional cases, positive ERG staining was found in morphologically benign glands, which were also immediately adjacent to or intermingled with ERG-positive cancerous glands. All other benign lesions distant from cancerous glands, including simple and partial atrophy, were negative for ERG. P63 was negative in all cancerous glands and positive in noncancerous lesions. The P63/ERG double immunostain combines the high sensitivity of P63 and the high specificity of ERG and may be potentially useful in the work-up of difficult prostate biopsies. The high specificity of ERG for the presence of cancer may have important implications for prostate biopsy interpretation and needs to be further validated in larger prospective studies.     

前列腺癌TSP-1的表达与血管新生的关系及意义

目的：探讨良性前列腺增生及前列腺癌中血管生成与血小板反应素-1（TSP-1）表达的相关性。方法：应用免疫组织化学方法检测32例良性前列腺增生和32例前列腺癌组织中TSP-1的表达及微血管密度（MVD）。结果：前列腺癌组织中TSP-1表达显著低于良性前列腺增生（P〈0．05）,MVD则明显升高（P〈0．05）;随着肿瘤分期的进展,浸润转移性癌中TSP-1的表达降低甚至缺失,而MVD却逐渐升高。结论：TSP-1在前列腺癌中呈低表达,与前列腺癌的血管新生相关;作为一种内源性血管新生抑制剂,它具有抑制肿瘤生长及血管新生的作用。