前列腺增生与前列腺癌中KLK11/TMPRSSmRNA表达比值

目的 以实时荧光定量PCR技术测定前列腺增生(BPH)与前列腺癌(PCa)组织标本KLK11/TMPRSS mRNA比值,探讨KLK11/TMPRSS比值在前列腺癌诊断的特异性意义.方法 通过实时荧光定量PCR对23例PCa、37例BPH及3例正常前列腺组织KLK11/TMPRSS的表达,比较其在PCa与BPH中组织定量的差异.结果 BPH与PCa组织KLK11/TMPRSS mRNA的定量表达值分别为2.264±0.460与5.905±0.780,差异有统计学意义(P＜0.05).结论 实时荧光RTPCR定量检测KLK11/TMPRSS mRNA为前列腺癌的诊断提供了可靠的辅助指标.     

前列腺癌与前列腺增生Ki-67/PCNA mRNA特异性表达比值的研究

目的 以实时荧光定量RT-PCR技术研究良性前列腺增生(BPH)与前列腺癌(PCa)组织标本Ki-67蛋白和组织核增殖抗原(PCNA)mRNA的比值,探讨此比值在PCa诊断中的特异性意义.方法 通过实时荧光定量RT-PCR检测63例PCa和37例BPH前列腺组织Ki-67/PCNAmRNA的表达,比较其在PCa与BPH组织中定量的差异.结果 BPH与PCa组织Ki-67/PCNA mRNA的定量表达值分别为2.264±0.460与5.905±0.780,差异有统计学意义(P＜0.05).结论 实时荧光定量RT-PCR检测Ki-67/PCNA mRNA为PCa的诊断提供了可靠的辅助指标.     

前列腺增生与前列腺癌特异性膜抗原mRNA的表达

目的以实时荧光定量聚合酶链反应(PCR)技术研究良性前列腺增生(BPH)与前列腺癌(Pca)组织标本PSMmRNA的表达,探讨前列腺特异性膜抗原(PSM)在前列腺癌诊断的特异性意义。方法通过实时荧光定量PCR对23例PCa、37例BPH及3例正常前列腺组织PSMmRNA的表达,比较其在3种组织定量的差异。结果BPH与PCa组织PSMmRNA的定量表达值分别为1.54±0.21与4.95±0.78,差异有统计学意义(P<0.05)。结论实时荧光RTPCR定量检测PSMmRNA为前列腺癌的诊断、治疗、预后监测等提供了更为可靠的辅助指标。     

实时荧光RT-PCR定量检测前列腺癌组织中AMACR基因表达

目的 建立实时荧光定量逆转录聚合酶链反应(FQ-RT-PCR)检测AMACR mRNA的方法学并检测其在前列腺癌(PCa)组织中的表达.方法 在AMACR基因的2、3外显子之间设计一对引物及MGB探针,将PCR扩增产物与pMD18-T载体连接,构建重组质粒作为定量检测的标准品,建立实时荧光定量RT-PCR方法,然后对32例PCa、60例良性前列腺增生(BPH)及34例其它肿瘤组织进行AMACR mRNA的定量检测.结果 重组质粒经PCR扩增及序列测定,表明克隆成功,该方法灵敏度高,线性范围为5～5×108copies/reaction.AMACR mRNA在PCa组织中表达量显著高于BPH组织(P<0.01)及其他类型肿瘤组(P<0.01).ROC曲线分析结果显示,曲线下面积(AUC-ROC)为0.890,AMACR mRNA诊断前列腺癌的敏感度和特异度分别为81.3%和86.7%.PCa组织中AMACR mRNA表达量及检测阳性率与不同临床分期和病理分级之间的差异尚不具有统计学意义(P值均>0.05).结论 实时荧光定量RT-PCR检测AMACR mRNA的方法具有敏感、特异、准确、重复性好等特点.AMACR mRNA在前列腺组织中的表达对PCa的诊断具有一定的临床应用价值.     

XAGE-1 b基因在良性前列腺增生症与前列腺癌中的表达及其意义

目的 观察XAGE-1b基因在良性前列腺增生症和前列腺癌中的表达,探讨其数值对良性前列腺增生症和前列腺癌各项指标的临床意义.方法 运用实时荧光定量聚合酶链反应(PCR)方法检测38例前列腺癌病理组织以及40例良性前列腺增生症组织XAGE-1b的基因表达水平.结果 XAGE-1b在前列腺癌组织中表达值为8.299 50±0.97116,在良性前列腺增生症组织中表达值为3.007 80±0.91600,差异有统计学意义(P＜0.05).XAGE-1b表达值随着Gleason评分、临床分期和肿瘤恶性程度升高而表达增强(P＜0.05).结论 XAGE-1b基因高表达值有助于前列腺癌的诊断和指导前列腺癌的恶性程度分期.

Abstract：

Objective To study the XAGE-1 b mRNA expression in prostate cancer (PCa) tissues and benign prostate hyperplasia (BPH) tissues,and explore the diagnostic values of XAGE-1 b mRNA expression level in PCa and BPH.Methods A sensitive,real-time quantitative polymerase chain reaction (PCR) assay was developed to compare the expression difference of XAGE-1b mRNA in PCa and BPH tissues,by testing 38 samples of PCa and 40 samples of BPH.Results The expression level of XAGE-1 b in PCa tissue was 8.299 50 ± 0.971 16,and 3.007 80 ± 0.916 00 in BPH tissue ( P＜0.05 ).The XAGE-1 b expression levels were increased with the increase of the Gleason score,clinical stage and malignant grade (P＜0.05).Conclusion The high expression of of XAGE-1 b mRNA can afford a reliable and helpful information for diagnosis of PCa and BPH,and PCa malignant grade.     

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。     

前列腺干细胞抗原在人前列腺癌组织中的表达及意义

目的　探讨前列腺干细胞抗原 (PSCA)在国人前列腺癌 (PCa)组织中表达的临床意义。　方法　采用免疫组织化学 (IHC)和核酸原位杂交 (ISH)方法检测 4 0例PCa、2 0例良性前列腺增生(BPH)和 2 0例前列腺上皮内瘤 (PIN)组织标本PSCA蛋白和mRNA表达 ,半定量法计算PSCA阳性表达细胞百分数和阳性表达强度 ,比较各组织间表达水平的差异及其与PCa分级、临床分期之间的关系。　结果　PCa、BPH、PIN组织PSCA中度阳性到强阳性表达分别为 85 % (34/ 4 0 )、2 0 % (4/ 2 0 )和35 % (7/ 2 0 ) ;PCa组织PSCA表达水平与BPH和PIN比较差异有统计学意义 (P <0 .0 5 ) ,BPH与PIN比较差异无统计学意义 (P >0 .0 5 ) ;PCa组织PSCA表达水平随Gleason评分及临床分期增加而升高。　结论　人PCa组织有PSCA蛋白质和mRNA的过表达 ,且与PCa病理分级、临床分期呈正相关 ,可能对PCa的诊断及判断预后有潜在价值。     

自噬相关基因Atg5与前列腺腺癌发生的相关性分析

目的:探讨自噬相关基因5(Atg5)与前列腺腺癌发生的相关性。方法:实时荧光定量PCR及免疫组化方法检测50例前列腺上皮内瘤变(PIN)、69例前列腺腺癌及30例良性前列腺增生(BPH)组织标本中Atg5的表达情况。结果:实时定量PCR结果显示,PIN及PCa组织中Atg5 mRNA的相对表达量显著高于BPH组织(5.270±0.230 vs 1.723±0.017,5.131±0.252 vs 1.723±0.017,P<0.01)。免疫组化结果显示Atg5在BPH、PIN及PCa组织中的阳性表达率分别为6.7%、94%及88.4%,BPH组织中Atg5的阳性表达率明显低于PIN及PCa,且差异具有统计学意义(P<0.001),PIN与PCa组别之间无显著差异(P>0.05)。Atg5的表达与前列腺腺癌的Gleason评分分级无显著相关性(P>0.05)。结论:前列腺组织中Atg5的高表达可能对前列腺腺癌的发生起着一定作用。     

原癌基因PIM-1检测在前列腺癌诊断中的临床意义

目的探讨原癌基PIM-1在前列腺癌诊断中的价值。方法实时荧光定量PCR法检测23例前列腺癌、37例良性前列腺增生及3例正常前列腺组织标本中PIM-1的表达,根据标准曲线法,比较3种组织中PIM-1定量的差异。结果前列腺癌、良性前列腺增生与正常前列腺组织PIM-1的定量表达值分别为4.45±0.63、2.57±0.74、1.05±0.04,3组间比较差异均有统计学意义(P<0.05)。结论实时荧光RT-PCR定量检测PIM-1有可能成为前列腺癌诊断的可靠辅助指标。     

实时荧光定量聚合酶链反应检测前列腺癌中Id1 mRNA的表达及意义

目的 检测Id1 mRNA在人前列腺癌（PCa）组织中的表达，探讨Id1 mRNA的表达与临床意义。方法 用SYBR Green Ⅰ嵌合荧光法进行实时逆转录-聚合酶链反应（RT—PCR），检测Id1 mRNA在人正常前列腺、前列腺良性增生（BPH）和PCa组织中的相对定量，分析Id1 mRNA的相对含量与PCa临床病理参数间关系。结果 Id1 mRNA在PCa组织中的含量明显高于BPH组织（P〈0．01）。Id1 mRNA表达量与PCa的Gleason评分有明显相关性（r=0．9995，P〈0．05）。Id1 mRNA表达量高者，2年内发生浸润和转移的机率提高。结论 PCa组织中Id1 mRNA表达明显增高，与肿瘤分化程度成负相关，Id1 mRNA的高表达从转录水平反映了其与前列腺癌病情进展的关系；Id1 mRNA的实时定量检测有助于从转录水平对前列腺癌进行早期诊断并初步指导预后。     

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。     

前列腺增生组织中白细胞介素-6受体基因表达水平的检测及其意义

应用逆转录聚合酶链反应方法定量检测了10例正常人前列腺、20例前列腺增生（BPH）组织及前列腺癌细胞系PC－3m中80ku白细胞介素－6受体（IL－6R）基因表达水平。结果发现正常前列腺组织中IL－6RmRNA呈低水平表达，而43PH组织中表达增强，明显高于正常组（P＜0．01），表明BPH发生与IL－6及其受体有关。此外，还发现55．0％（11／20）BPH组织中IL－6RmRNA含量高于前列腺癌细胞系PC－3m，提示IL－6所介导的融合毒素可能成为治疗BPH新的有效途径。     

High expression of PSM-E correlated with tumor grade in prostate cancer: a new alternatively spliced variant of prostate-specific membrane antigen

BACKGROUND: Prostate-specific membrane antigen (PSMA) overexpressed in prostate cancer (PCa) has been targeted for therapy and diagnosis of PCa. In the current study, PSMA cDNA was cloned from PCa tissue by RT-PCR. After sequencing, a new spliced variant of PSMA (PSM-E) was discovered and its specificity in PCa was evaluated. METHODS: PSM-E and PSMA mRNA were measured in LNCaP, PC-3 and prostate or nonprostatic malignancies. Following transfection of PC-3 with PSM-E cDNA in the pcDNA3.0 vector, PSM-E expression was measured by immunofluorescence and Western-blot. PSM-E and PSMA mRNA levels were quantified by a real-time PCR assay in normal prostate (n = 7), benign prostatic hyperplasia (BPH) (n = 22) and PCa (n = 41). The correlation between their levels and tumor grade was analyzed. RESULTS: PSM-E cDNA is identical to PSMA except for a 97-nucleotide region and a 93-nucleotide region. PSM-E and PSMA mRNA were detected in PCa and LNCaP, not in PC-3; PSMA could be detected in some nonprostatic tumors whereas PSM-E not. The expression of PSM-E protein was detected in transfected cells. Significant difference of PSM-E mRNA levels was observed among normal prostate, BPH and PCa (P < 0.001), and PSM-E levels increased with increasing Gleason score (r = 0.514, P < 0.001). PSMA mRNA levels were higher in BPH and PCa than in normal prostate (P < 0.001), but no difference between BPH and PCa, no significant correlation was observed between PSMA levels and Gleason score (r = 0.229, P = 0.057). CONCLUSIONS: PSM-E may be a potential prognostic indicator for PCa progression and may be a new target antigen for therapy of PCa.     

Dysregulation of Dkk-3 expression in benign and malignant prostatic tissue

BACKGROUND: The Dickkopf (Dkk) family comprises four members Dkk-1, -2, -3, and -4. Dkk-3, the most divergent family member, unlike the others does not modulate Wnt signaling. Dkk-3 is proposed to function as a secreted tumor suppressor since it is downregulated in a number of cancer cells and prostate cancer tissue and thus may be a promising candidate molecule for therapeutic interference. METHODS: The in situ tissue localization of Dkk-3 protein in normal prostate (NP), benign prostatic hyperplasia (BPH), and prostate carcinoma (PCa) was investigated by immunohistochemistry (IHC)/immunofluorescence. In addition, biological function of Dkk-3 in terms of proliferation and viability was evaluated in primary prostate basal epithelial cells (PrEC), stromal cells (PrSC), and established human PCa cell lines by treatment with recombinant protein or by overexpression. RESULTS: Stimulation with purified recombinant protein and overexpression of Dkk-3 did not significantly alter in vitro cell proliferation in any primary or tumor cell line evaluated. Dkk-3 was expressed in both the basal and secretory epithelium of NP. In BPH expression was restricted to defined basal cells and was absent in tumor cells of high grade PCa. In contrast to normal prostatic tissue, Dkk-3 was upregulated in subglandular blood vessels of BPH and in the reactive stroma of PCa tissue. CONCLUSIONS: Our results indicate that Dkk-3 expression in the normal epithelium of the prostate is lost during benign and malignant transformation and differentiation processes. The loss of expression seems to be counterbalanced by upregulation of Dkk-3 expression in the blood vessels of the remodeled tissue.     

Upregulation of erythropoietin receptor in human prostate carcinoma and high-grade prostatic intraepithelial neoplasia

The aim of the present study was to investigate the differential expression of erythropoietin receptor (EPOR) in prostate carcinoma (PCa), high-grade prostatic intraepithelial neoplasia (PIN), prostatic hyperplasia (BPH) lesions and normal prostatic tissues by immunohistochemistry; and to test the hypothesis that upregulation of EPOR is a specific event for prostate carcinogenesis. An immunohistochemical analysis of EPOR was performed on 30 PCa, 50 BPH with/without inflammation lesions and 30 normal prostatic tissue samples. EPOR staining was quantitated and classified into normal expression and overexpression. Totally 16 high-grade PIN lesions were found in this study. Overexpression of EPOR was shown only in PCa and high-grade PIN. Statistical analysis demonstrated that higher median EPOR staining score of PCa and high-grade PIN in comparison with BPH (P < 0.05) and higher median EPOR staining score of PCa compared with high-grade PIN (P < 0.05). Our data demonstrate that upregulation of EPOR is not uncommon for PCa and upregulated EPOR in high-grade PIN suggests upregulation of EPOR is an early event for prostate carcinogenesis. The role of upregulated EPOR and possibly enhanced EPOR signaling in prostate carcinogenesis warrants further studying.