FOXN2在前列腺癌组织的表达及对前列腺癌细胞生物学的影响

探讨双头框蛋白N2（FOXN2）在前列腺癌（PCa）组织中的表达以及对PCa细胞生物学的影响。方法 选取宜宾市第二人民医院50例接受手术治疗的PCa患者的标本及癌旁组织,通过RT-qPCR实验和Western blot实验分别检测PCa组织和癌旁组织中的FOXN2 mRNA及蛋白的表达水平,并分析PCa组织中FOXN2与患者临床病理特征的相关性。通过RT-qPCR实验检测正常前列腺细胞RWPE-1、PCa细胞PC-3、DU145、LNCaP中FOXN2 mRNA的表达水平。选取PC-3细胞为研究对象,分为FOXN2过表达组、空载体对照组以及对照组,分别通过MTT实验、流式细胞实验、Transwell实验以及Western blot实验检测各组细胞增殖、凋亡、迁移和侵袭情况以及相关蛋白Bax、CyclinD1、MMP-2的表达量。结果 与癌旁组织相比,FOXN2的mRNA和蛋白表达水平显著降低（P<0.05）,与TCGA数据库中结果一致。FOXN2低表达与PCa患者淋巴转移、TNM分期以及Gleason评分有关（P=0.003、0.005、0.002）。与人正常前列腺细胞RWPE-1相比,FOXN2在PCa细胞PC-3、DU145、LNCaP中均呈现低表达（P<0.05）,其中PC-3细胞中表达量最低。与对照组和空载体对照组相比,FOXN2过表达组的PC-3细胞在作用24 、48 、72 h后增殖能力显著下降（F=290.400、57.735、113.014,P<0.05）,CyclinD1蛋白表达水平显著下降（P<0.05）;PC-3细胞的凋亡率显著升高（P<0.05）,Bax蛋白表达水平显著升高（P<0.05）;PC-3细胞的迁移和侵袭能力显著下降（P<0.05）,MMP-2蛋白表达水平显著下降（P<0.05）。结论 FOXN2在PCa组织及细胞中低表达,过表达FOXN2可抑制PCa细胞的增殖、迁移和侵袭,并促进细胞凋亡。     

前列腺特异性膜抗原表达的临床意义

目的 探讨前列腺特异性膜抗原(PSMA)在前列腺癌(PCa)组织中的表达及与肿瘤病理分级之间的关系。方法 采用免疫组化ABC法，用PSMA单克隆抗体对前列腺不同病变组织及非前列腺肿瘤组织石蜡包埋切片进行染色。其中PCa 70例，前列腺上皮肉瘤21例，良性前列腺增生20例．其他肿瘤组织标本30例。结果 PSMA在97％前列腺癌、100％前列腺上皮内瘤、80％BPH组织中呈不同程度的阳性表达，且在PCa组织中呈明显高表达，非前列腺肿瘤组织染色均呈阴性。组织 PSMA表达与PCa组织学分级之间存在负相关性。结论 PSMA具有良好的组织器官特异性，能够判断PCa顶后，在PCa的免疫治疗方面具有应用前景。     

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。     

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.     

前列腺癌组织中TRIB1基因和蛋白异常表达与患者不良预后的相关性

目的研究Tribbles同源蛋白1(TRIB1)在良性前列腺增生和前列腺癌中的表达,并分析其表达水平与患者临床特征及预后的关系。方法在公共基因芯片数据库(GEO)中下载前列腺样本中的相关基因芯片数据,分析TRIB1在前列腺样本中mRNA的表达水平。收集临床手术切除或者穿刺活检的前列腺癌和前列腺增生组织,通过免疫组化检测前列腺癌、前列腺增生组织中TRIB1蛋白的表达。分析TRIB1蛋白和基因的表达水平与患者临床特征及预后的关系。结果前列腺癌患者组织中TRIB1蛋白表达显著上调(P0.01),TRIB1蛋白表达上调与前列腺癌Gleason评分(P0.01)、病理分期(P=0.02)相关,基因芯片数据提示TRIB1基因表达上调与前列腺癌Gleason评分(P=0.02)、病理分期(P=0.01)、无生化复发生存率(BCR-free survival)(P=0.047)相关。结论 TRIB1在前列腺癌组织中表达上调,前列腺组织中检测TRIB1可能有助于判断前列腺癌分化程度并评估预后。     

凋亡抑制基因XIAP在前列腺癌细胞中的表达及与临床病理特征的关系

目的 :研究XIAP基因在前列腺癌细胞系和前列腺癌组织的表达情况 ,及其与前列腺癌临床病理特征的关系。　方法 :应用RT PCR检测前列腺癌组织、正常前列腺组织和前列腺癌细胞株PC 3,DU 14 5 ,LNCaP细胞XIAP基因的表达 ,并通过免疫组化SP法检测 5 6例前列腺癌组织标本XIAP蛋白的表达情况。　结果 :XIAP基因在前列腺癌组织和前列腺癌细胞株PC 3,DU 14 5 ,LNCaP细胞高表达 ,正常前列腺组织无表达。在前列腺癌组织和癌旁组织中 ,XIAP蛋白阳性检出率分别为 5 3.6 % (30 / 5 6 )和 2 1.5 % (12 / 5 6 ) (P <0 .0 1) ;不同分期、分级组XIAP阳性检出率相比差异无显著性 (P >0 .0 5 )。　结论 :凋亡抑制基因XIAP与前列腺癌相关 ,在前列腺癌发生过程中具有重要的作用 ,有可能成为前列腺癌治疗的靶标。     

低氧诱导因子1α及血管内皮生长因子在前列腺癌中的表达及意义

目的:观察低氧诱导因子1α(H IF-1α)及血管内皮生长因子(VEGF)在前列腺癌(PCa)中的表达及意义。方法:32例PCa患者,根据G leason评分,将≥7分者设为高G leason评分组(n=12),<7分者为低G leason评分组(n=20)。良性前列腺增生(BPH)16例,BPH伴高级别前列腺上皮内瘤(PIN)15例,正常前列腺组织(NP)12例。采用免疫组化染色CD34观察各组组织中微血管密度(MVD)及H IF-1α、VEGF的表达情况。结果:PCa、PIN中H IF-1α阳性表达率分别为62.5%、60.0%,较BPH(6.3%)及NP(0)高,差异有统计学意义(P<0.05)。PCa、PIN中VEGF阳性表达率分别为78.1%、73.3%,较BPH(18.7%)及NP(8.3%)高,差异亦有统计学意义(P<0.05)。PCa的MVD为66.9±18.0,明显高于BPH(28.3±6.9)及NP(15.3±2.9)(P<0.05)。高G leason评分组H IF-1α、VEGF阳性率及MVD值均高于低G leason评分组,差异均有显著性(P<0.05)。结论:H IF-1α及VEGF过表达是PCa形成的早期事件,与PCa密切相关。     

前列腺癌与良性前列腺增生中NFKBIA及SREBF2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NFKBIA及SREBF2 mRNA相对值,探讨NFKBIA及SREBF2比值在前列腺癌诊断中的特异性意义.方法 通过实时荧光定量PCR检测63例PCa、37例BPH及3例正常前列腺组织NFKBIA及SREBF2的表达,比较其在PCa与BPH组织中定量的差异.结果 与BPH比较,PCa组织NFKBIA及SREBF2 mRNA的定量表达值分别下调0.520和上调2.547,差异有统计学意义(P＜0.05).结论 实时荧光逆转录(RT)-PER定量检测NFKBIA及SREBF2 mRNA为前列腺癌的诊断提供了参考.     

Down-regulation of macrophage inhibitory cytokine-1/prostate derived factor in benign prostatic hyperplasia

BACKGROUND: Macrophage inhibitory cytokine-1 (MIC-1) is a member of transforming growth factor-beta/bone morphogenetic protein (BMP) superfamily. Despite its potential role in prostatic regulation, little is known about its biological activity. METHODS: Expression profiling using 42K Affymetrix HuGeneFL array was conducted to compare symptomatic benign prostatatic hyperplasia (BPH), histological BPH without symptoms, and normal prostate samples from donors. MIC-1 gene expression was analyzed by RT-PCR in pure culture of prostate epithelial and stromal cells, and prostate cancer cells, LNCaP, PC-3, DU-145. Influence of androgens on MIC-1 expression in LNCaP cells was analyzed by Northern blot. Enhancement of promoter activity of MIC-1 by androgens was examined using reporter assays. RESULTS: In contrast to normal prostates, MIC-1 gene was down-regulated in BPH samples with symptoms and histological BPH obtained from cystoprostatectomy specimens (P < 0.005 and P < 0.01, respectively). Expression level of MIC-1 in androgen-sensitive LNCaP cells was high and enhanced by androgens, whereas in the androgen-insensitive PC-3 and DU-145 cells the expression level was low. An 11 kb promoter region of MIC-1 gene was identified to be 6- to 12-fold activated by androgens. CONCLUSIONS: Down-regulation of MIC-1 may play a role in the development of BPH. MIC-1 is positively regulated by androgens, but other regulatory factors remain unclear.     

Cell-Surface labeling and internalization by a fluorescent inhibitor of prostate-specific membrane antigen

BACKGROUND: [corrected] Prostate-specific membrane antigen (PSMA) remains an attractive target for imaging and therapeutic applications for prostate cancer. Recent efforts have been made to conjugate inhibitors of PSMA with imaging agents. Compared to antibodies, small-molecule inhibitors of PSMA possess apparent advantages for in vivo applications. To date, there are no reports on the cellular fate of such constructs once bound the extracellular domain of PSMA. The present study was focused on precisely defining the binding specificity, time-dependent internalization, cellular localization, and retention of inhibitor conjugates targeted to PSMA on LNCaP cells. A novel fluorescent inhibitor was prepared as a model to examine these processes. METHODS: Fluorescence microscopy of LNCaP and PC-3 cell lines was used to monitor the specificity, time-dependent internalization, cellular localization, and retention of a fluorescent PSMA inhibitor. RESULTS: Fluorescent inhibitor 2 was found to be a potent inhibitor (IC50 = 0.35 nM) of purified PSMA. Its high affinity for PSMA on living cells was confirmed by antibody blocking and competitive binding experiments. Specificity for LNCaP cells was demonstrated as no labeling by 2 was observed for negative control PC-3 cells. Internalization of 2 by viable LNCaP cells was detected after 30 min incubation at 37 degrees C, followed by accumulation in the perinuclear endosomes. It was noted that internalized fluorescent inhibitor can be retained within endosomes for up to 150 min without loss of signal. CONCLUSIONS: Our results suggest that potent, small-molecule inhibitors of PSMA can be utilized as carriers for targeted delivery for prostate cancer for future imaging and therapeutic applications.     

环氧化酶2在不同前列腺癌细胞系中的表达

目的:研究环氧化酶2(COX-2)在不同前列腺癌细胞系中的表达,探讨COX-2在前列腺癌侵袭进展及转移潜能获得机制中的可能作用。方法:应用Western印迹及RT-PCR鉴定LNCaP及其亚细胞系C4-2和AR-CaP亚细胞系IF11、IA8,以及PC-3细胞中COX-2的表达情况,并初步分析其在不同特性前列腺癌细胞系转移侵袭过程中的作用。结果:Western印迹结果显示:COX-2蛋白在PC-3细胞中表达相对较高,在IF11、IA8、LNCaP和C4-2细胞中表达缺失,差异具有统计学意义(P<0.05)。COX-2mRNA表达结果同蛋白一致。结论:不同来源、不同转移潜能的前列腺癌细胞株中COX-2表达存在差异。高表达COX-2可能在PC-3细胞高侵袭转移潜能获得方面起着一定作用,而与其他细胞系转移作用无关。     

组织激肽释放酶基因7在前列腺癌组织中的表达

目的 探讨组织激肽释放酶基因7(KLK7)在不同前列腺组织中的表达情况.方法运用逆转录聚合酶链反应法检测正常前列腺(5例)、良性前列腺增生(BPH)及BPH细胞株(BPH1,13例)、前列腺癌及前列腺癌细胞株(8例)的上皮细胞中KLK7mRNA表达水平;蛋白质印迹法检测不同前列腺组织上皮细胞中KLK7蛋白表达水平;免疫组化分析正常前列腺(20例)、BPH(50例)、前列腺癌(103例)组织中KLK表达水平.根据染色强度分为4个等级(-,+,++,+++)进行半定量分析,染色强度++及+++者判定为阳性.结果 正常组、BPH组和前列腺癌组KLK7 mRNA表达相对值分别为0.59、0.52、0.02,组间比较差异有统计学意义(F=13.03,P＜0.01),前列腺癌上皮中KLK7 mRNA表达下调(P＜0.01),正常前列腺和BPH上皮中KLK7 mRNA表达差异无统计学意义(P＞0.05).KLK7蛋白在正常前列腺、增生前列腺、DU145、LNCaP、PC3、22RV1、BPH细胞株中表达水平相对值分别为0.22、0.40、0.01、0.05、0、0.03、0.14.免疫组化染色结果 显示正常前列腺组织、BPH组织、前列腺癌中KLK7蛋白表达阳性率分别为65.0%(13/20)、76.0%(38/50)、17.5%(18/103),前列腺癌组与前2组比较差异均有统计学意义(P＜0.01),前2组间比较差异无统计学意义(P＞0.05).结论 KLK7在前列腺癌组织中表达下调,提示KLK7在前列癌的发生和进展中可能起一定作用.     

miR-18a在前列腺癌患者血清中的表达及其临床意义

目的探讨前列腺癌患者microRNA-18a(miR-18a)在血清中的表达及其诊断价值。方法采用实时荧光定量PCR(qRT-PCR)检测60例前列腺癌患者(PCa)、30例良性前列腺增生患者(BPH)和20例健康对照者(HC)血清miR-18a的表达水平。分析miR-18a表达水平与前列腺癌Gleason分级、TNM分期及PSA的关系。通过分析受试者工作特征(ROC)曲线判断miR-18a表达水平在前列腺癌诊断中的灵敏度和特异度。结果PCa患者血清miR-18a的表达水平分别明显高于BPH组和HC组(P<0.01);miR-18a与Gleason评分、肿瘤分期有关(P<0.01);miR-18a与PSA之间呈正相关(r=0.701,P=0.000);miR-18a的ROC曲线下面积(AUC)为0.928(95%CI:0.880~0.976,P=0.000),敏感度为84.3%,特异度为75.8%。结论miR-18a在PCa患者血清中表达水平明显增高,对于PCa的诊断有潜在的临床参考价值。     

Prostate cancer antigen-1 as a potential novel marker for prostate cancer

Aim： To examine the expression of prostate cancer antigen-1 （PCA-1） in prostate cancer （PCa） and to validate it as a potential marker for diagnosis of PCa. Methods： In situ hybridization analysis of PCA-1 mRNA expression was performed on 40 benign prostate hyperplasia （BPH）, 16 high-grade prostatic intraepithelial neoplasm （HG-PIN）, 74 PCa and 34 other malignant carcinoma specimens. The level of PCA- 1 expression was semiquanfitatively scored by assessing both the percentage and intensity of PCA- 1 positive staining cells in the specimens. We then compared the PCA-1 expression between BPH, HG-PIN and PCa and evaluated the correlation of PCA-1 expression level with clinical parameters of PCa. Results： PCA-1 mRNA was expressed in the majority of both PCa and HG-PIN specimens but not in BPH and other malignant carcinoma. The expression level of PCA-1 increased along with a high Gleason score （P 〈 0.05）, and was unrelated to other clinical parameters of PCa （all P 〉 0.05）. Conclusion： The data suggest that PCA-1 might be a novel diagnostic marker for PCa, and that increased PCA-1 expression might denote more aggressive variants of PCa.     

Increased expression and differential phosphorylation of stathmin may promote prostate cancer progression

BACKGROUND: Proteins which regulate normal development may promote tumorigenesis, tumor progression, or metastasis through dysregulation of these functions. We postulate that proteins, which regulate prostate growth also promote prostate cancer (PCa) progression. METHODS: Two Dimensional Gel Electrophoresis was utilized to compare patterns of protein expression in 12T-7f prostates (LPB-Tag mouse model for PCa) during tumor development and progression with those of normal developing and adult wild type CD-1 prostates. Stathmin expression and phosphorylation patterns were analyzed in mouse and human PCa cell lines as well as in human PCa tissue arrays. RESULTS: Stathmin was identified by two-dimensional gel electrophoresis and mass spectrometry. Stathmin levels increase early during normal mouse prostate development and again during prostate tumor development and progression. In human prostate adenocarcinoma, stathmin increases in Gleason pattern 5. Further, stathmin is differentially phosphorylated in androgen-dependent LNCaP cells compared to androgen-independent PC-3 and DU145 cells. This differential phosphorylation is modulated by androgen and anti-androgen treatment. CONCLUSION: Stathmin expression is highest when the prostate is undergoing morphogenesis or tumorigenesis and these processes may be regulated through differential phosphorylation. Furthermore, modulation of stathmin phosphorylation may correlate with the development of androgen-independent PCa.     

Increased expression of anterior gradient-2 is significantly associated with poor survival of prostate cancer patients

Anterior gradient-2 (AGR2) expression was examined in a series of prostate cell lines and in an archival set of prostate tissues. The relative levels of AGR2 expression in the malignant cell lines PC-3 and PC-3M were, respectively, 5.3+/-0.1 and 3.8+/-0.2 times that detected in the benign cell line PNT-2. Immunohistochemical staining in 106 cases showed that amongst seven normal cases, one (14.3%) was unstained, five (71.4%) stained weakly positive and one (14.3%) stained moderately positive. Amongst 34 benign prostate hyperplastic (BPH) cases, 12 (35.3%) were unstained, 18 (52.9%) stained weakly positive and four (11.8%) stained moderately positive. Amongst 65 carcinomas, three (4.6%) were unstained, 14 (21.5%) stained weakly positive, 19 (29.2%) stained moderately positive and 29 (44.9%) stained strongly positive. AGR2 expression in carcinomas was significantly higher than that in BPH (chi(2)-test, P<0.001). Kaplan-Meier survival analysis showed that increased AGR2 expression was significantly (log rank test, P=0.007) associated with reduced patient-survival time. Increased joint Gleason score (GS) was significantly (log rank test, P=0.001) associated with poor patient survival. However, neither prostate specific antigen (PSA) level, nor androgen receptor (AR) index, was significantly associated with patient-survival time. Increased AGR2 expression was significantly correlated with high GS (two-sided Fisher's exact test, P<0.001) and PSA levels (Mann-Whitney U-test, P=0.047), but not significantly related to the level of AR (Mann-Whitney U-test, P=0.286). These results suggest that increased AGR2 expression is a valuable prognostic factor to predict the clinical outcome of the prostate cancer patients.     

前列腺癌和良性前列腺增生磁共振灌注加权成像的应用价值

目的 探讨磁共振灌注加权成像对前列腺癌(PCa)和良性前列腺增生(BPH)的诊断和鉴别诊断价值.方法 收集行磁共振灌注加权成像检查的PCa患者23例、BPH患者22例及正常对照组8例,分别比较其前列腺灌注参数的差异,包括相对负性强化积分(rNEI)、相对平均达峰时间(rMTE)和相对最大下降斜率(rMSD).结果 外周带PCa组rNEI值高于BPH组外周带(t=11.64,P＜0.001)和正常外周带(t=11.79,P＜0.001);外周带PCa组rMTE值低于BPH组外周带(￡=3.50,P=0.002)和正常外周带(t=3.78,P=0.001).中央带PCa组rNEI值高于BPH组中央带(t=6.99,P＜0.001)和正常中央带(t=10.32,P＜0.001);BPH组中央带rMSD值高于中央带PCa组(t=2.30,P=0.027)和正常中央带(t=3.15.P=0.001).应用受试者工作特征曲线分析,rNEI对PCa具有诊断意义,选择最佳诊断阈值为6.44.敏感性为91.3%(21/23)、特异性为81.8%(18/22).结论 磁共振灌注加权成像的rNEI可作为鉴别PCa与BPH的有效指标.     

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.