Differential localization of MT1-MMP in human prostate cancer tissue: role of IGF-1R in MT1-MMP expression

BACKGROUND: MT1-MMP is a metalloproteinase involved in prostate cancer metastasis. The IGF-1R is a tyrosine kinase receptor involved with tumor progression and metastasis. The purpose of this investigation was to examine MT1-MMP and IGF-1R expression and localization in prostate cancer tissues and explore the role of IGF-1R in regulating MT1-MMP in prostate cancer cell lines. METHODS: Immunohistochemistry was utilized to study MT1-MMP and IGF-1R expression in human prostate tissues. IGF-1R regulation of MT1-MMP expression was determined by gene promoter analysis, quantitative RT-PCR and Western blot analysis following pharmacological inhibition of the receptor in PC-3N cells and treatment of LNCaP cells with androgen and IGF-1. RESULTS: MT1-MMP expression was high in the apical regions of the luminal cells in PIN and prostate cancer and less intense in the basalateral regions of benign tissues. IGF-1R was expressed primarily in the basal cells of normal glands and highly expressed in prostate cancer. Inhibition of IGF-1R in PC-3N cells decreased MT1-MMP expression and treatment of LNCaP cells with a synthetic androgen and IGF-1 increased MT1-MMP expression. CONCLUSIONS: These data demonstrate that MT1-MMP is highly expressed in the apical cytoplasmic regions of the luminal cells in PIN and prostate cancer when compared to basalateral cytoplasmic membrane staining in benign glands. Additionally, we demonstrate that IGF-1R is highly expressed in human prostate carcinoma. These findings suggest that MT1-MMP localization and IGF-1R expression in prostate carcinoma could be predictive biomarkers for aggressive disease and support IGF-1R as a promising therapeutic target to decrease processes of prostate cancer metastasis.     

Glucose transporter-1 gene expression is associated with pancreatic cancer invasiveness and MMP-2 activity

BACKGROUND: Overexpression of the facilitative glucose transporter-1 (GLUT-1) has been observed for a wide range of human cancers, with the degree of overexpression generally being inversely correlated with prognosis. We tested the effects of modulating GLUT-1 expression on pancreatic cancer cellular invasiveness. METHODS: GLUT-1 expression in MIAPaCa-2, PANC-1, BXPC-3, and CAPAN-2 cells was assayed using Western blotting. Cells were stably transfected with a GLUT-1 expression vector or a GLUT-1 RNA interference vector to alter GLUT-1 expression. Matrix metalloproteinase-2 (MMP-2) activity and expression were assayed using zymography and Western blotting, respectively. In vitro cellular invasiveness was assayed using Matrigel Boyden chambers, and in vivo metastatic potential was assessed using a nude mouse xenograft model. RESULTS: Variable baseline GLUT-1 expression levels were detected among the cell lines. Forced overexpression of GLUT-1 induced increases in MMP-2 expression and activity and in cellular invasiveness. GLUT-1 silencing induced reductions in MMP-2 expression and activity, cellular invasiveness, and metastatic potential in vivo. CONCLUSION: GLUT-1 promotes pancreatic cellular invasiveness. The therapeutic implications of this finding warrant further study.     

Chromosome 8q arm overexpression is associated with worse prostate cancer prognosis

ObjectiveChromosome 8q arm (chr8q) is the most amplified chromosomal segment in advanced metastatic castration-resistant prostate cancer after chXq12. These regions harbor important oncogenes driving prostate cancer progression, including MYC that plays a role in various hallmarks of cancer, including cell cycle progression and immune surveillance. Herein we characterize the co-expression patterns of chr8q genes and their clinical utility in more than 7,000 radical prostatectomy samples.Materials and methodsCopy Number alterations of 336 genes on chr8q21 to chr8q24 were extracted from 2 primary prostate cancer cohorts (TCGA, n = 492; MSK-primary, n = 856) and 3 metastatic prostate cancer cohorts (MSK-met, N = 432; MSK-mCSPC, N = 424; SU2CPNAS, n = 444) from cBioPortal. Expression data for the 336 genes was extracted from 6,135 radical prostatectomy samples from Decipher GRID registry. For survival analysis, patients were grouped into top 10% and top 25% by band expression and were compared with the remaining cohort. Hazard ratios were calculated using Cox proportional hazards models.ResultsGenes on chr8q were highly co-amplified and co-expressed. Copy number alterations and overexpression of chr8q genes in primary disease were associated with higher Gleason scores, increased risk of metastases, and increased prostate cancer specific mortality. Additionally, our data demonstrated high expression of MYC alone was not associated with differences in metastases free survival while high expression of other chr8q bands was associated with decreased metastases free survival. By combining chr8q data with an established genomic classifier like Decipher, we were able to develop a new model that was better at predicting metastases than Decipher alone.ConclusionsOur findings highlight the clinical utility of chr8q data, which can be used to improve prognostication and risk prediction in localized prostate cancer.     

GREB1 tissue expression is associated with organ-confined prostate cancer

ObjectiveBy reason of its heterogeneous behavior, it is difficult to determine the prognosis of many prostate cancer cases. Patients with the same clinicopathologic conditions may present varying clinical findings and rates of progression. We determined the role of new genes as potential molecular markers for prostate cancer prognosis.Materials and methodsWe performed a microarray analysis of two pools of patients with prostate cancer divided according to their clinicopathologic characteristics. After that, we validated these results by testing the genes with most different expressions between the two pools using the quantitative real time polymerase chain reaction method. We analyzed gene expression in 33 patients with localized prostate cancer according to prostate specific antigen (PSA), pathologic stage, Gleason score, and biochemical recurrence. For statistical analysis we used the Mann-Whitney Test.ResultsThe microarray analysis revealed that 4,147 genes presented a different expression between the two pools. Among them, 3 genes, TMEFF2, GREB1, and TH1L, were at least 13-times overexpressed, and 1 gene, IGH3, which was at least 5times under-expressed in pool 1 (good prognosis) compared with pool 2 (bad prognosis), were selected for analysis. After the validation tests, GREB1 was significantly more overexpressed among patients with stage T2 compared with T3 (P = 0.020). The expressions of other 3 genes did not present significant differences according to the clinicopathological variables.ConclusionsTissue expression of GREB1 is associated with organ-confined prostate cancer and may constitute a gene associated with a favorable prognosis.     

Caveolin-1 overexpression is associated with aggressive prostate cancer recurrence

BACKGROUND: Caveolin-1 protein suppresses apoptotic cell death in prostate cancer. The objectives of this study were to investigate the association of Caveolin-1 expression with established features of prostate cancer as well as overall and aggressive disease recurrence in patients treated with radical prostatectomy (RP). METHODS: Caveolin-1 immunostaining was performed on a tissue microarray containing prostatectomy specimen cores from 232 consecutive patients treated with RP for clinically localized prostatic adenocarcinoma. Caveolin-1 over-expression was defined as more than 50% of cells staining positively for Caveolin-1. Patients were categorized as having features of aggressive disease recurrence if they had a positive metastatic work-up, post-recurrence PSA doubling time less than 10 months, and/or failure to respond to local salvage radiation therapy. RESULTS: Seventy patients (30.2%) exhibited over-expression of Caveolin-1. Caveolin-1 over-expression was associated with higher pathologic Gleason sum (P=0.038) and higher pre-operative PSA level (P=0.024). Patients with Caveolin-1 over-expression were at increased risk of PSA recurrence after surgery (P=0.023) in univariate but not in standard post-operative multivariate analysis. However, patients with Caveolin-1 over-expression were at increased risk of aggressive prostate cancer recurrence in both univariate and multivariate analysis (P<0.001 and P=0.001, respectively). CONCLUSIONS: Over-expression of Caveolin-1 was associated with established features of prostate cancer and aggressive PSA recurrence. Caveolin-1 might help identify patients at high risk of developing aggressive prostate cancer recurrence, thus allowing selection of patients who might benefit from early systemic therapeutic intervention.     

纳米材料碘油混悬液栓塞后兔VX2肿瘤组织MT1-MMP表达改变的意义

目的 观察应用羟基磷灰石纳米粒子和超液化碘油混悬液栓塞后,兔VX2肿瘤组织膜型基质金属蛋白酶-1(MT1-MMP)表达变化的意义.方法 60只接种VX2肿瘤细胞的荷瘤兔随机分为3组:肿瘤组、超液化碘油组、羟基磷灰石纳米超液化碘油混悬液组.通过胃十二指肠动脉插管分别给予:生理盐水(1 ml/只)、超液化碘油(0.3 ml/kg)、纳米超液化碘油混悬液(0.3 ml/kg).术后3 d CT检查确认插管成功.两周后,应用免疫组织化学三步法(S-P)测定肿瘤组织MT1-MMP的表达定位,以及治疗干预后表达量的改变.RT-PCR检测MT1-MMP mRNA的表达差异,Western blot法分析MT1-MMP蛋白表达变化.结果 MT1-MMP在肿瘤细胞膜和肿瘤组织间质都有表达.肿瘤组与碘油组、纳米碘油组的阳性表达相比,差异有统计学意义(P＜0.05),而碘油组和纳米碘油组则无明显差异(P＞0.05).Western blot法检测各组蛋白量的表达也支持此结果.RT-PCR检测3组mRNA表达值的对比无明显差异(P＞0.05).结论 MT1-MMP主要表达于肿瘤细胞膜和间质.超液化碘油和(或)羟基磷灰石纳米粒子栓塞后,肿瘤组织及间质的MT1-MMP的表达上调,可能是造成栓塞后肿瘤转移复发率高的分子机制之一.     

8p22 loss concurrent with 8c gain is associated with poor outcome in prostate cancer

OBJECTIVES: A critical issue in the management of prostate cancer is the ability to distinguish patients at risk of disease recurrence. The aim of this study was to determine whether specific physical alterations of chromosome 8 may be associated with disease recurrence and poor outcome using postoperative prostate-specific antigen (PSA) values as surrogate end points. METHODS: To test this hypothesis, we examined paired normal and tumor radical prostatectomy tissues from 25 patients with prostate cancer for chromosome 8 alterations using dual fluorescence in situ hybridization with a fluorescein-labeled 8p22-specific (8p) cosmid probe and a rhodamine-labeled 8-centromere-specific (8c) probe. The probes were enumerated in 200 nuclei per tissue. RESULTS: Of the 25 tumors examined, 22 demonstrated distinct classes of genetic alterations, or nuclear types, including disomy for 8p and 8c (1 tumor), loss of 8p and disomy for 8c (10 tumors), or loss of 8p concurrent with gain of 8c (11 tumors). The presence of even a small population of tumor nuclei characterized by the loss of 8p concurrent with the gain of 8c was correlated with poor tumor grade (P = 0.009), preoperative PSA values 11 ng/mL or higher (P = 0.022), high tumor stage (P = 0.086), and detectable, rising postoperative PSA values (P = 0.086). These observations are consistent with the hypothesis that a gain of chromosome 8 is associated with poor outcome in prostate cancer. CONCLUSIONS: 8p loss concurrent with 8c gain may successfully predict disease recurrence and poor clinical outcome before the observation of detectable postoperative PSA values in patients with prostate cancer.     

Circulating syndecan-1 is associated with chemotherapy-resistance in castration-resistant prostate cancer

Objectives

Docetaxel chemotherapy is a standard treatment for castration-resistant prostate cancer (CRPC). Rapidly expanding treatment options for CRPC provide reasonable alternatives for those who are resistant to docetaxel. Therefore, prediction of docetaxel resistance has become of great clinical importance. Syndecan-1 (SDC1) has been currently shown to be involved in chemotherapy resistance in various malignancies including prostate cancer. The predicting value of serum SDC1 level has not been evaluated yet.

Matrix metalloproteinase-2 polymorphism is associated with prognosis in prostate cancer

ObjectiveProstate cancer (PCa) is the most frequent tumor in males in Brazil. Single nucleotide polymorphisms (SNP) have been demonstrated in the promoter region of matrix metalloproteinases (MMPs) genes and have been associated with development and progression of some cancers. In this study, our aim was to investigate a possible relation between polymorphism of the promoter region of the MMP2 gene and classical prognostic parameters in prostate cancer.Materials and methodsGenomic DNA was extracted using conventional protocols. The DNA sequence containing the polymorphic site was amplified by real-time polymerase chain reaction, using fluorescent probes (TaqMan).ResultsIn patients with tumors of a higher stage (pT3), a polymorphic allele in the MMP2 gene was more frequent (P = 0.026) than in patients with lower tumor stage. A polymorphic allele in the MMP2 gene was more frequent in Gleason ≥ 7 than in Gleason ≤ 6 (P = 0.042).ConclusionsWe conclude that MMP2 polymorphism can be used together with pathological stage and Gleason score to identify patients with worse prognosis. Our results illustrate the potential use of MMP2 SNP as a molecular marker for prostate cancer.     

DLC2和RhoA蛋白在原发性肝癌组织中的表达及其临床意义

目的:探讨肝癌缺失基因2(DLC2)和RhoA蛋白在肝癌组织中的表达及其与预后的关系。方法:采用Western印迹法和免疫组化技术检测128例原发性肝癌组织中DLC2蛋白和RhoA蛋白的表达;分析DLC2蛋白的表达与肝癌临床病理特征的关系及DLC2蛋白对原发性肝癌预后的价值。结果:DLC2蛋白在HCC组织中的表达较癌旁组织低,差异具有统计学意义(P<0.05)。DLC2蛋白在细胞分化程度差的肝癌组织中表达降低(P<0.05)。同时,DLC2蛋白的低表达与RhoA蛋白的高表达呈正相关。DLC2蛋白表达低的原发性肝癌患者预后更差。结论:原发性肝癌组织中DLC2蛋白的低表达与较差的细胞分化程度,RhoA蛋白的高表达相关,DLC2缺失表达的原发性肝癌患者预后较差。     

Self-reported acne is not associated with prostate cancer

ObjectiveSome studies have suggested an inverse association between acne vulgaris and the acne-related bacterium Propionibacterium acnes and prostate cancer (PCa). Self-reported acne might be an easily obtainable marker to identify men at relatively low risk of PCa and might be incorporated into PCa risk calculators. This study aimed to evaluate the association between self-reported acne and PCa in a large case-referent study.Methods and materialsThe case group comprised 942 patients with PCa recruited from a population-based cancer registry in 2003 to 2006, 647 of whom met the criteria for aggressive PCa. The referents (n = 2,062) were a random sample of the male general population. All subjects completed a questionnaire on risk factors for cancer, including questions about acne. Odds ratios (ORs) and 95% confidence interval (CI) were calculated using multivariable logistic regression for PCa and aggressive PCa as separate end points, while adjusting for age and family history of PCa.ResultsA history of acne was reported by 320 cases (33.9%) and 739 referents (35.8%). Self-reported acne was significantly associated neither with PCa (adjusted OR = 0.95, 95% CI: 0.80–1.12) nor with aggressive PCa (adjusted OR = 0.97, 95% CI: 0.80–1.18).ConclusionSelf-reported acne is not suitable as a marker to identify men at low risk of aggressive PCa.     

Plasma concentrations of metalloproteinases (MMP-1, MMP-3) and their inhibitors in patients with prostate cancer

Summary Matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) are involved in important processes of tumor invasion and metastasis. In the study presented, matrix metalloproteinase 1 (MMP1) and 3 (MMP3), the tissue inhibitor of metalloproteinase 1 (TIMP1) and the complex MMP1/TIMP1 were measured by ELISA tests specific for these proteins in blood plasma. These components have been investigated in prostate cancer patients (PCa) with metastases (n = 18; T2, 3, 4 pN1, 2M1), prostate cancer patients without metastases (n = 29; T2, 3 pN0M0), patients with benign prostate hyperplasia (BPH; n = 29) and in healthy men (n = 35). Mean values of MMP1 and of the complex MMP1/TIMP1 were not different among the four groups. The mean values of MMP3 and especially TIMP1 were significantly higher in prostate cancer patients with metastases compared with controls, BPH patients and prostate cancer patients without metastases. Ten of these 18 patients had TIMP1 levels higher than the upper reference limit. TIMP1 concentrations correlate to the tumor stage but not to the tumor grade. These results indicate, that TIMP1 could be an potential marker for metastases in prostate cancer patients.      

前列腺液中IL-1β、IL-8、TNF-α、PGE2浓度变化及临床意义

目的 探讨慢性前列腺炎患者前列腺液中IL-1β、IL-8、TNF-α、PGE2的浓度变化与慢性前列腺炎类型、发病机制、症状和前列腺液白细胞计数的关系.方法 以Meares-Stamey法行尿液、前列腺按摩液(EPS)细菌培养,结合前列腺液常规检查和NIH-CPSI评分,将100例慢性前列腺炎患者分为细菌组(Ⅱ型11例)、非细菌组(Ⅲa型66例)、前列腺痛组(Ⅲb型23例).放射免疫法测定患者和20例正常对照者前列腺液中IL-1β、IL-8、TNF-α、PGE2含量,比较结果并进行相关分析.结果 Ⅱ型前列腺炎患者EPS中IL-1β、IL-8、TNF-α、PGE2检测值明显高于Ⅲa型、Ⅲb型及对照组(P＜0.01),Ⅲa型明显高于Ⅲb型及对照组(P＜0.05),Ⅲb型与对照组相比差异无统计学意义(P＞0.05).前列腺炎患者IL-1β与TNF-α、PGE2水平成正相关(r=0.563,r=0.826;P＜0.01).EPS中白细胞计数与IL-8水平成正相关(r=0.547,P＜0.01),与IL-1β、TNF-α、PGE2水平无相关性;IL-1β、PGE2水平与NIH-CPSI疼痛不适症状评分成正相关(r=0.506,P＜0.01);IL-1β、IL-8、TNF-α、PGE2水平与NIH-CPSI评分无相关性(P＞0.05).结论 慢性前列腺炎患者前列腺液IL-1β、IL-8、TNF-α、PGE2表达增高,并参与前列腺的炎症反应,其水平可作为慢性前列腺炎的诊断依据之一,对慢性前列腺炎分型可能有一定临床意义.     

联合检测泛素连接酶Cullin1和基质金属蛋白酶MT4-MMP在乳腺癌组织中的表达及临床意义

目的探究泛素连接酶Cullin1和基质金属蛋白酶MT4-MMP在不同类型乳腺癌组织中表达情况及临床意义。 方法2015年1月至8月,选择徐州市肿瘤医院80例乳腺癌组织及其对应的癌旁正常乳腺组织,应用免疫组织化学法和Western blotting法分别检测其中Cullin1和MT4-MMP的表达情况,并分析其相关性。 结果Cullin1、MT4-MMP蛋白在乳腺癌组织中的表达均高于癌旁正常乳腺组织,差异均有统计学意义（77.5% vs 28.8%,67.5% vs 17.5%,χ²=38.174、40.921,均P<0.01）;在乳腺癌组织中,Cullin1及MT4-MMP蛋白的表达呈正相关（P<0.05）。Cullin1及MT4-MMP在乳腺癌中的蛋白表达在分子分型、腋窝淋巴结是否转移及TNM分期组间差异有统计学意义（P<0.05）;在患者年龄、肿瘤大小、肿瘤部位、是否复发转移组间差异无统计学意义。 结论Cullin1及MT4-MMP可能参与了乳腺癌的形成及发展过程,且两者在此过程中有一定协同作用。在分型差、分期晚的乳腺癌中,Cullin1及MT4-MMP更高的表达状态提示其可能成为新的乳腺癌恶性程度指标。     

Flap endonuclease 1 is overexpressed in prostate cancer and is associated with a high Gleason score

OBJECTIVE: To investigate the expression and potential clinical usefulness of structure-specific flap endonuclease 1 (FEN-1) in human primary prostate cancer using tissue microarray technology, as FEN-1 was recently identified to be overexpressed in CL1.1, the most aggressive clone generated from the hormone-refractory prostate cancer cell line CL1. MATERIALS AND METHODS: Immunohistochemistry was performed on tissue microarrays constructed from paraffin-embedded specimens of primary prostate cancer from 246 patients who had had a radical prostatectomy. Prostatic intraepithelial neoplasia (PIN), benign prostatic hyperplasia (BPH) and normal prostate epithelium were represented on the array. FEN-1 nuclear expression was scored based on the percentage of target cells staining positively, and correlated with Gleason score, preoperative prostate-specific antigen (PSA) level and pathological stage. The time to PSA recurrence was also analysed. RESULTS: The mean expression of FEN-1 was significantly higher in cancer (36.7%) than in normal (13.2%), BPH (4.5%) and PIN (15.4%) specimens (P < 0.001). FEN-1 expression was significantly correlated with Gleason score (ó = 0.23, P = 0.002). A higher preoperative serum PSA level (P = 0.015), Gleason score > or = 7 (P < 0.001), seminal vesicle invasion (P < 0.001) and capsular involvement (P = 0.004) were associated with PSA recurrence, whereas FEN-1 expression was not. In a multivariate analysis, only Gleason score > or = 7 (P < 0.001), seminal vesicle invasion (P = 0.005) and capsular involvement (P = 0.009) were retained as independent predictors for PSA recurrence. CONCLUSIONS: FEN-1 is overexpressed in prostate cancer compared with matched normal prostate, and its expression increases with tumour dedifferentiation, as shown by increasing Gleason score. These results suggest that FEN-1 might be a potential marker for selecting patients at high risk, and a potential target for prostate cancer diagnosis and therapy.     

Human kallikrein-2 gene polymorphism is associated with the occurrence of prostate cancer

PURPOSE: Human glandular kallikrein, which is encoded by the human kallikrein-2 (KLK2) gene, is significantly associated with the occurrence of prostate cancer (PCa). We tested the association between a functional C748T polymorphism of the KLK2 gene and the occurrence of PCa. MATERIALS AND METHODS: Peripheral venous blood samples were obtained from 254 patients with PCa and 168 controls with benign prostatic hyperplasia. All control subjects had normal serum prostate specific antigen and proved to be free from malignancy on pathological examination of resected prostatic tissues. Serum prostate specific antigen, testosterone, Gleason score, clinical and pathological stage, tumor and prostate volume of the patients were investigated. The KLK2 gene polymorphism was determined by the polymerase chain reaction based restriction fragment length polymorphism method. RESULTS: The PCa group had a younger mean age +/- SEM (73.0 +/- 0.5 vs 74.7 +/- 0.5 years, p = 0.010) and higher C allele frequency (82.1% vs 74.7%, p = 0.010) than the control group. The frequency of the CC, CT and TT genotypes was 65.7%, 32.7% and 1.6% in patients with PCa, and 56.0%, 37.5% and 6.5%, respectively, in controls (p = 0.010). C allele carriers (CC and CT genotypes) were at significantly higher risk for PCa than TT homozygous subjects (p = 0.002). CC homozygous subjects were at significantly higher risk for PCa than T allele carriers (CT and TT genotypes) (p = 0.043). The PCa subgroup of patients with pathologically proved, localized PCa also had a higher frequency of the C allele (87.5% vs 74.7%, p = 0.026) and CC genotypes (78.7% vs 56.0%, p = 0.019) than controls. CONCLUSIONS: Our results suggest that the C allele of the functional C748T polymorphism of KLK2 may increase the risk of PCa.