TMSG-1在人前列腺癌细胞株与前列腺癌组织中的表达及临床意义

目的 探讨肿瘤转移抑制基因-1(TMSG-1,亦称LASS2)在人不同转移潜能前列腺癌细胞株中与前列腺癌组织中的表达及其临床意义.方法 采用实时荧光定量聚合酶链反应(PCR)及细胞爬片免疫荧光组织化学方法,检测TMSG-1在人不同转移潜能前列腺癌细胞株低转移潜能(PC-3M-2134)和高转移潜能(PC-3M-IE8)中的表达.并采用免疫组织化学方法检测TMSG-1在人前列腺增生及前列腺癌组织中的表达,同时探讨其与临床病理特征之间的关系.结果 TMSG-1在PC-3M-284细胞株中的mRNA及蛋白表达(2.70±0.30、75.26±2.68)均明显高于在PC-3M-IE8细胞株中的表达(1.10±0.20、38.08±1.84),差异有统计学意义(P＜0.05).通过免疫组织化学观察表明TMSG-1在前列腺增生及前列腺癌组织中均有表达,但在前列腺增生中的阳性表达率(32/40)明显高于在前列腺癌中的阳性表达率(21/60).两者差异有统计学意义(P＜0.05).并且TMSG-1在前列腺癌组织中的表达与年龄、Gleason分级、淋巴结转移及TNM分期密切相关(P＜0.05),而与肿瘤的大小无明显相关.结论 TMSG-1在低转移潜能前列腺癌细胞株中的mRNA及蛋白表达明显高于在高转移潜能前列腺癌细胞株中的表达,证明它是一种肿瘤转移抑制基因.TMSG-1在人前列腺增生与前列腺癌组织中的表达之间差异有统计学意义,并且TMSG-1在前列腺癌中的表达与年龄、Gleason分级、淋巴结转移及TNM分期密切相关.

Abstract：

Objective To investigate the expression of tumor metastasis suppressor gene 1 (TMSG-1 as well LASS2) in different prostate cancer cell lines and prostate cancer tissues and its clinical significance. Methods Sixty patients with prostate cancer had undergone surgery between 2008 and 2010.Forty patients with prostatic hyperplasia were chosen. Immunofluorescence histochemistry was used to study the distribution of TMSG-1 in cells, immunohistochemistry was used to observe the difference in TMSG-1 expression between prostatic hyperplasia and prostate cancer tissues, and the relationship between the TMSG-1 expression and clinicopathological features in prostate cancer tissues was analyzed. Results The level of TMSG-1 mRNA in PC-3M-2B4 cell line with low metastatic potentiality (2. 70 ±0. 30) was higher than in PC-3M-IE8 cell line (1. 10 ±0. 20). Immunofluorescence histochemistry revealed that most of the collected prostate cancers and prostatic hyperplasia tissues expressed TMSG-1 in cytoplasma, and nuclei were stained in a few of prostate cancer tissues. The average fluorescence intensity of TMSG-1 in PC-3M-2B4 cells (75. 26 ±2. 68) was obviously higher than in PC-3M-IE8 cells (38. 08 ± 1. 84). There was obviously different expression of TMSG-1 between prostate cancers (21/60) and prostatic hyperplasia ( 32/40 ) ( P ＜0. 05 ) . The TMSG-1 levels in prostate cancer tissue were significantly correlated with ages,Gleason grade, lymph node metastasis and tumor, nodes, metastasis (TNM) staging (P ＜0. 05) , but not with the size of tumor. Conclusion The expression level of TMSG-1 mRNA and protein in prostate cancer cell lines with low metastatic potentials significantly higher than in prostate carcinoma cell lines with high metastatic potentials, which proves that TMSG-1 is a tumor metastasis suppressor gene. From the difference in the TMSG-1 expression between human prostatic hyperplasia and prostate cancer tissues and the correlation with age, Gleason grade, lymph node metastasis and TNM stage in prostate cancers, we infer that TMSG-1 is an important prognostic indicator in judging prostate cancer cell growth, progression and metastasis.     

P_(53)蛋白和增殖细胞核抗原在膀胱癌中的表达及意义

采用免疫组化法对69例膀胱移行细胞癌中P_(53)蛋白及增殖细胞核抗原(PCNA)进行检测。发现膀胱癌中P_(53)蛋白的过度表达与病理分级无关,而与临床分期及预后有关。PCNA-LI与病理分极、临床分期及预后均相关,随着肿瘤分级、分期的增高,PCNA-LI呈明显上升趋势;P_(53)蛋白过度表达或PCNA高表达组术后5年生存率明显低于P_(53)蛋白非过度表达或PCNA低表达组。结果表明:P_(53)蛋白的过度表达在膀胱癌发生、发展中起着一定作用,PCNA是判定膀胱癌恶性程度及预后的重要指标,同时发现膀胱癌中P_(53)蛋白的过度表达与PCNA-LI相关。     

p53和VEGF在前列腺癌组织中的表达及临床意义

目的研究p53和血管内皮生长因子（vascular endothelial growthfactor,VEGF）在同一前列腺癌（prostate cancer,Pca）组织中的表达及与Pca临床参数的关系,探讨肿瘤血管形成及调节机制。方法应用免疫组织化学LDP法检测46例Pca组织及20例良性前列腺增生症（benign prostatic hyperplasia,BPH）组织中p53蛋白及VEGF的表达。结果Pca组织中p53与VEGF阳性表达率分别为41.30%（19/46）和71.74%（33/46）,均明显高于BPH组（P〈0.05）;p53与VEGF表达呈明显正相关（P〈0.05）;二者在Pca组织中的表达水平与其病理分级和临床分期均呈正相关（P〈0.05）;p53和VEGF阳性表达的肿瘤复发迅速、易转移,p53阴性而VEGF阳性表达较阴性表达者预后差。结论p53和VEGF与Pca组织学分级？恶性程度和预后密切相关,是检测Pca的较好分子标志物;Pca是典型的血管依赖性病变,p53可能通过p53-VEGF调节旁路途径促进Pca的肿瘤血管形成,联合检测p53和VEGF的表达可作为判断Pca生物学行为及预后的重要指标。     

MTS_1基因产物P_(16)蛋白在肾癌中的表达及意义

采用免疫组化法对42例肾癌、20例癌旁正常肾组织MTS_1基因产物P_(16)蛋白进行检测。结果显示肾癌P_(15)蛋白表达阳性率明显高于癌旁正常肾组织(P<0.01),P_(16)表达相关于肾癌病理分级、临床分级及预后。结果提示:MTS_1基因与肾癌的发生发展有关,P_(16)蛋白表达缺失是肾癌恶性度高及晚期表现。P_(16)蛋白表达可作为判断肾癌预后的指标。     

Ets-1在前列腺癌组织中的表达及其临床意义

目的 探讨原癌基因Ets-1在前列腺组织中的表达及其与前列腺癌组织学分级的关系.方法 利用免疫组化法检测Ets-1在4例正常前列腺组织,12例良性前列腺增生组织和53例前列腺癌组织中的表达.结果 Ets-1蛋白的阳性表达在前列腺癌细胞的胞浆及胞核内均可见,79%的前列腺癌组织中Ets-1呈阳性表达,而在良性前列腺组织细胞中无表达或仅有微弱表达,差异有显著性意义(P<0.05).在前列腺癌组织中,Gleason评分高危组Ets-1阳性表达率明显高于Gleason中危组和低危组(P<0.05).结论 与良性前列腺组织对照相比,Ets-1有肿瘤特异性,其表达与前列腺癌的组织学分级相关,可作为前列腺癌病变程度及临床预后判断的一个分子标志.     

组织芯片检测前列腺癌中BRG1表达的研究

目的:探讨前列腺癌组织中BRG1基因的表达情况及临床病理意义。方法:运用组织芯片技术、免疫组化EnV ision法检测37例前列腺癌[其中9例中分化腺癌(G leason评分5~7分),28例低分化腺癌(G leason评分8~10分)]、13例前列腺上皮内瘤、14例良性前列腺增生组织中BRG1的表达和分布情况。结果:BRG1在前列腺癌、前列腺上皮内瘤和良性前列腺增生组织中阳性表达率分别为81.08%(30/37)、38.46%(5/13)、14.28%(2/14)。就BRG1阳性表达率而言,前列腺癌组织与前列腺上皮内瘤、前列腺增生组织比较差异均有显著性(P<0.05),前列腺上皮内瘤与前列腺增生组织比较、中分化腺癌与低分化腺癌比较差异均无显著性(P>0.05)。结论:BRG1蛋白在前列腺癌中高表达,对前列腺癌的发生、发展可能起重要作用。组织芯片技术具有高信息量、简捷、快速、高效、成本低、重复性好等优点,在病理学领域中具有重要的实际意义和广阔的应用前景。     

Yap1蛋白在前列腺癌中的表达及其临床病理学意义

目的 探讨Yap1蛋白在前列腺癌中的表达及临床病理学意义.方法 应用免疫组织化学方法检测Yap1在35例前列腺癌组织和16例良性前列腺增生组织中的表达,用平均吸光度值(积分吸光度/面积,IA/area)代表阳性细胞的表达水平;并探讨Yap1与前列腺癌临床病理参数之间的关系.结果 Yap1在良性前列腺增生组的表达水平(IA/area =0.1867 ±0.0753)明显高于前列腺癌组(IA/area=0.0782 ±0.0614),两组差异有统计学意义(P＜0.01,F=16.942);Yap1蛋白主要表达在细胞质.Yap1蛋白表达程度与前列腺癌的Gleason评分无明显相关(P＞0.05),但Yapl在Gleason 7分组(IA/area =0.0548±0.0441)的表达强度高于Gleason 9分组(IA/area=0.1296±0.0957) (P ＜0.05).结论 前列腺癌中Yap1蛋白低表达可能与前列腺的发生、发展有关.     

TRAIL受体在前列腺癌中的表达

目的　探讨肿瘤坏死因子相关诱导凋亡配体 (TRAIL)受体在前列腺癌组织中的表达及其与前列腺癌恶性度的关系。方法　采用免疫组织化学方法检测前列腺癌组织及良性前列腺增生组织中DR4、DR5及DcR1的表达。结果　前列腺癌组织DcR1表达水平显著低于良性前列腺增生组织 (P <0 .0 1) ,DR4、DR5的表达水平两者无显著性差异。前列腺癌组织中DR4、DR5及DcR1的表达程度与前列腺癌组织的病理分级和临床分期无关 (P >0 .0 5 )。结论　TRAIL基因受体DcR1在前列腺癌凋亡机制中可能发挥重要作用     

生育酚结合蛋白在前列腺癌组织的表达及意义

目的探讨生育酚结合蛋白(α-TAP)在良恶性前列腺组织的表达差异及意义。方法构建TAP-N端的质粒并转化到大肠杆菌BL21-Condon Plus,以1 mmol/L IPTG诱导3 h表达蛋白,用Ni-NTA琼脂糖柱纯化TAP蛋白并免疫兔生产抗体。将pGEM-T-easy质粒酶切线性化,37℃体外转录120 min合成地高辛标记的TAP-RNA探针。免疫组织化学和原位杂交法测定TAP在分别40例良性前列腺增生(BPH)与前列腺癌(PCa)的表达。结果制备到毫克量的TAP-N端蛋白、特异性TAP抗体和RNA探针。TAP主要表达于前列腺上皮,很少位于基质成分。BPH组TAP表达阳性率(37例,92．5%)比PCa组高(7例,17．5%),两组差异有统计学意义(P＜0．05)。无TAP表达的33例PCa患者术前PSA水平(12．5±3．3)μg/L、Gleason评分(7．3±1．4)分均高于其余7例TAP阳性患者(P＜0．05),后者PSA为(6．7±2．6)μg/L、Gleason评分为(4．6±2．1)分。TAP阴性的PCa患者术后出现复发时间(32．5±6．2)个月小于TAP阳性患者(78．5±12．6)个月(P＜0．05)。结论TAP在前列腺癌的表达比在良性前列腺增生组织明显下调,无TAP表达的前列腺癌患者术前PSA和Gleason评分高于TAP阳性患者。     

前列腺癌增殖细胞核抗原表达的意义

目的：研究前列腺癌增殖细胞核抗原的意义表达。方法：应用HE染色、免疫组化染色、光学显微镜观察对20例前列腺癌和15例正常前列腺组织中增殖细胞核抗原（PCNA）表达进行检测。结果：前列腺癌中增殖细胞核抗原的表达明显高于正常前列腺组织。结论：PCNA是判断肿瘤细胞增殖程度的重要指标，对估计前列腺癌的分化、转移及预后具有重要参考价值。     

胃癌及其癌旁病变中p16、nm23-H_l基因蛋白的表达及意义

目的:应用免疫组化方法探讨p16蛋白在胃癌发生中的作用.方法:应用免疫组化方法对57例胃癌切除标本进行p16蛋白和nm23-H_1基因产物的表达检测.结果:57例胃癌中p16蛋白表达阳性率为40/57(70.2%),显著低于癌旁正常胃粘膜(P<0.01).胃癌高分化组的阳性率显著高于低分化组(P     

前列腺癌组织中PSCA在蛋白质和mRNA水平表达相关性分析及其意义

目的:探讨人前列腺癌组织中前列腺干细胞抗原(PSCA)蛋白和mRNA的表达状况及其相关性.方法:采用免疫组化和原位杂交方法同步检测48例前列腺癌组织中PSCA蛋白和mRNA的表达,并进一步比较其表达水平之间的相关性.结果:免疫组化和原位杂交检测显示:42例(87.5%)的PSCA蛋白阳染区域所见与其mRNA阳染区域相同,其中的37例的蛋白和mRNA表达水平(评分值)均一致,其余11例的PSCA蛋白表达水平与其mRNA表达水平比较,均有不同程度的降低.结论:PSCA蛋白和mRNA在人前列腺癌组织中的表达水平具有很高的一致性,PSCA蛋白表达与PSCA基因的转录密切相关.     

P53蛋白和增殖细胞核抗原在肾癌中的表达及意义

采用免疫组化法对５３例肾癌组织中Ｐ５３蛋白及增殖细胞核抗原（ＰＣＮＡ）进行检测。发现肾癌组织Ｐ５３蛋白表达与细胞类型、分级、分期及预后均无关。肾癌ＰＣＮＡ表达与细胞类型、分级、分期及预后均相关，透明细胞癌与颗粒细胞癌差异无显著性，梭形细胞癌呈高表达；随肿瘤分级增高，ＰＣＮＡ表达呈明显上升趋势；肿瘤高分期组（Ⅲ、Ⅳ）的ＰＣＮＡ表达明显高于低分期组（Ⅰ，Ⅱ）；ＰＣＮＡ高表达组（３、４级）术后５年生存率明显低于低表达组（１、２级）。结果表明：Ｐ５３蛋白表达在肾癌发生中并不起主导作用或只是多种因素之一，ＰＣＮＡ是判定肾癌恶性度及预后的客观指标，同时发现肾癌Ｐ５３蛋白与ＰＣＮＡ表达间无相关。     

胃癌组织中p16,p53蛋白的表达及其临床意义

为探讨ｐ１６，ｐ５３蛋白在胃癌发生、发展中的作用及其临床意义。应用免疫组化方法，对６０例胃癌、１８例胃良性疾病及１５例胃正常组织中ｐ１６，ｐ５３蛋白的表达情况进行检测。结果显示：胃癌组织中ｐ１６蛋白的阳性率（２５％）低于胃良性疾病（６１．１１％）及胃正常组织中的阳性率（６３．３３％），有显著性差异（Ｐ＜０．０５）；而胃癌组织中ｐ５３蛋白的阳性率（６１．６７％）高于胃良性疾病（１１．１１％）及胃正常组织（０），差异非常显著（Ｐ＜０．００１）。ｐ１６，ｐ５３蛋白的表达与胃癌的组织学分化、临床分期及淋巴结转移密切相关，与患者年龄、性别、肿瘤位置无关；ｐ１６蛋白表达还与肿瘤大小有关。研究结果表明，ｐ１６蛋白的失表达发生在胃癌发生、发展的晚期阶段，ｐ１６，ｐ５３蛋白可作为判别胃的良、恶性疾病及胃癌患者预后的一个指标。     

Androgen Receptor Mediated Growth of Prostate (Cancer)

The understanding of the mode of action of androgens requires insight in the cell biological architecture of the prostate. In the prostate secretory acini, morphologically two cell layers can be discriminated; i.e. the basal and the luminal compartment. The stem cells are thought to be located in the basal compartment. The stem cells have the unique capacity of self-renewal, providing the full repertoire to develop the differentiated ductal system via transient proliferating/amplifying (TP/A) intermediate stem cells. These are in fact early and late progenitors for the exocrine/secretory and neuro-endocrine cells. Exocrine differentiation occurs in the majority of the luminal compartment and is identified by the expression of the androgen receptor (AR), keratin 18 (K18) and prostate specific antigen (PSA). Neuro-endocrine cells are dispersed in the prostate epithelium and express K5 and typical neuro-endocrine markers such as chromogranin A and/or bombesin. The exocrine lineage of differentiation is critically dependent on androgens and thus androgen deprivation therapy will result in declining numbers of secretory/exocrine cells, while the number of stem cells and TP/A intermediate stem cells remains stable. This implies that stem cells and TP/A intermediate stem cells are for their renewal androgen independent, although the TP/A intermediate stem cells are androgen sensitive for expanding the epithelial compartment.Recently, these TP/A intermediate stem cells gained attention because they are thought to play an important role in normal prostate growth as well as in neoplastic transformation. Current hypotheses suggest that from the TP/A cell population, the cancer stem cell develops. Hypothetically, the more committed the cancer stem cell is, the more sensitive the tumour might be for androgen ablation, which could explain why some patients are long-term hormone therapy responders, while others are intrinsically androgen independent. In conclusion, typing of the transformed TP/A intermediate stem cells could enable to discriminate long-term hormone responders from non-responders and could help individualisation of prostate cancer therapy in the future.     

己糖激酶1在肾癌中的异常表达

目的探讨己糖激酶1(hexokinase 1,HK1)基因在人肾癌和癌旁组织中差异表达。方法提取47例肾癌及其癌旁组织的RNA和蛋白,荧光定量实时聚合酶链反应(real-time quantitative polymerase chain reaction,QPCR)检测HK1的表达水平,应用免疫组化方法检测HK1蛋白在肾癌及相应癌旁组织的表达情况。结果癌组织△CT值为4.43±1.54,癌旁组织的△CT值为5.59±1.70,二者有统计学差异(t=-4.97,P=0.00)。37例(78.7%,37/47)肾癌组织样本中HK1表达量高于癌旁组织。免疫组化结果显示,HK1蛋白表达于胞浆内,在肾癌组织中的表达量明显高于相应的正常组织。结论 HK1高表达于肾癌组织,可能在肿瘤发生发展的过程中发挥重要作用,该基因有望成为指导肾癌诊断及治疗的新靶点。     

Clinical Outcomes in Cyclin-dependent Kinase 12 Mutant Advanced Prostate Cancer

BackgroundCyclin-dependent kinase 12 (CDK12) loss occurs in 3–7% of metastatic prostate cancer patients and is characterized by a genomic instability signature, but the clinical implications of CDK12 loss are not well established.ObjectiveTo determine the clinical course of patients with CDK12 mutant advanced prostate cancer compared with other genomic subtypes.Design, setting, and participantsA retrospective analysis of data from three academic medical centers, including 317 patients with advanced prostate cancer and prior next-generation sequencing from tumor tissue (n = 172) or circulating tumor DNA (n = 145), was performed. Forty-six patients had CDK12 mutations; 34 had biallelic CDK12 loss (79%).Outcome measurements and statistical analysisPatients were stratified by mutation status (CDK12, homologous recombination deficiency [HRD; BRCA1/2 and ATM], TP53, and other cohort). The Kaplan-Meier method was used to evaluate time to event outcomes: time to development of metastatic disease, time to development of castration resistance, and time to prostate-specific antigen (PSA) progression after first-line androgen receptor pathway inhibitor (ARPI) therapy in a patient subset.Results and limitationsThe median follow-up was 66.6 mo. Patients with CDK12 mutant prostate cancer exhibited shorter time to metastasis (median = 34.9 mo, p = 0.004) and development of castration-resistant disease (median = 32.7 mo, p < 0.001), compared with other genomic subtypes, with shorter time to PSA progression on first-line ARPI treatment of metastatic castration-resistant disease (median = 3.6 mo, p = 0.0219). CDK12 mutant patients did not have overall shorter time on treatment compared with other mutation subgroups, and CDK12 status did not demonstrate statistical significance in multivariate analysis. Limitations include variable center-dependent practice patterns and heterogeneity due to combining tumor and liquid biopsy data.ConclusionsOur data suggest that advanced prostate cancers harboring CDK12 mutations display aggressive clinical behavior, underscoring the need to fully delineate the molecular and clinical characteristics, and appropriate therapeutic approaches for distinct subtypes of advanced prostate cancers.Patient summaryIn this report, we evaluate the clinical characteristics and outcomes of patients with prostate cancer and CDK12 mutation in their tumors. These patients seem to have more aggressive disease, with more high-grade Gleason ≥8 cancers and shorter time to developing metastatic cancer. Cases of advanced CDK12-mutated prostate cancer may warrant consideration of therapy intensification or combination approaches.     

Prostate Cancer (PCa) Risk Variants and Risk of Fatal PCa in the National Cancer Institute Breast and Prostate Cancer Cohort Consortium

Background

Screening and diagnosis of prostate cancer (PCa) is hampered by an inability to predict who has the potential to develop fatal disease and who has indolent cancer. Studies have identified multiple genetic risk loci for PCa incidence, but it is unknown whether they could be used as biomarkers for PCa-specific mortality (PCSM).

Objective

To examine the association of 47 established PCa risk single-nucleotide polymorphisms (SNPs) with PCSM.

Design, setting, and participants

We included 10 487 men who had PCa and 11 024 controls, with a median follow-up of 8.3 yr, during which 1053 PCa deaths occurred.

Outcome measurements and statistical analysis

The main outcome was PCSM. The risk allele was defined as the allele associated with an increased risk for PCa in the literature. We used Cox proportional hazards regression to calculate the hazard ratios of each SNP with time to progression to PCSM after diagnosis. We also used logistic regression to calculate odds ratios for each risk SNP, comparing fatal PCa cases to controls.

Results and limitations

Among the cases, we found that 8 of the 47 SNPs were significantly associated (p < 0.05) with time to PCSM. The risk allele of rs11672691 (intergenic) was associated with an increased risk for PCSM, while 7 SNPs had risk alleles inversely associated (rs13385191 [C2orf43], rs17021918 [PDLIM5], rs10486567 [JAZF1], rs6465657 [LMTK2], rs7127900 (intergenic), rs2735839 [KLK3], rs10993994 [MSMB], rs13385191 [C2orf43]). In the case-control analysis, 22 SNPs were associated (p < 0.05) with the risk of fatal PCa, but most did not differentiate between fatal and nonfatal PCa. Rs11672691 and rs10993994 were associated with both fatal and nonfatal PCa, while rs6465657, rs7127900, rs2735839, and rs13385191 were associated with nonfatal PCa only.

Conclusions

Eight established risk loci were associated with progression to PCSM after diagnosis. Twenty-two SNPs were associated with fatal PCa incidence, but most did not differentiate between fatal and nonfatal PCa. The relatively small magnitudes of the associations do not translate well into risk prediction, but these findings merit further follow-up, because they may yield important clues about the complex biology of fatal PCa.

Patient summary

In this report, we assessed whether established PCa risk variants could predict PCSM. We found eight risk variants associated with PCSM: One predicted an increased risk of PCSM, while seven were associated with decreased risk. Larger studies that focus on fatal PCa are needed to identify more markers that could aid prediction.