Aberrant neuropeptide Y and macrophage inhibitory cytokine-1 expression are early events in prostate cancer development and are associated with poor prognosis.

Studies to elucidate dysregulated gene expression patterns in premalignant prostate lesions have identified several candidate genes with the potential to be targeted to prevent the development and progression of prostate cancer and act as biomarkers of early disease. Herein, we explored the importance of two proteins, neuropeptide Y (NPY) and macrophage inhibitory cytokine-1 (MIC-1), as biomarkers of preinvasive prostate disease and investigated the relationship of expression to biochemical recurrence following treatment for localized prostate cancer. NPY and MIC-1 protein expression was determined by immunohistochemistry on tissue microarrays containing 1,626 cores of benign, low-grade prostatic intraepithelial neoplasia (PIN), high-grade PIN (HGPIN), and prostate cancer tissue from 243 radical prostatectomy patients. Both NPY and MIC-1 showed higher proportional immunostaining in HGPIN and prostate cancer compared with benign epithelium (P < 0.0001). NPY and MIC-1 immunostaining was higher in low-grade PIN compared with other benign tissues (both P < 0.0001) and was equivalent to immunostaining in HGPIN. NPY immunostaining of prostate cancer was independently associated with relapse, after adjusting for traditional prognostic factors, as a categorical variable in 20% intervals (P = 0.0449-0.0103) and as a continuous variable (P = 0.0017). Low MIC-1 immunostaining (20% categories) was associated with pathologic stage >2C after adjusting for predictors of pathologic stage (P = 0.3894-0.0176). This is the first study to show that altered NPY and MIC-1 expression are significantly associated with prostate cancer progression and suggests that these molecules be developed further as biomarkers in the management of prostate disease.     

MIC-1、VEGF和P53蛋白表达与直肠癌临床病理及预后的关系

背景与目的：巨噬细胞是重要的免疫效应细胞,对肿瘤的发生、发展发挥着重要的调控作用.本研究探讨巨噬细胞抑制细胞因子-1（MIC-1）、血管内皮生长因子（VEGF）、P53蛋白的表达与直肠癌临床病理特征及预后的相关性.方法：用免疫组化法检测73例直肠癌中MIC-1、VEGF和P53的表达,并与临床病理因素及预后进行相关性分析.结果：MIC-1表达与VEGF、P53表达呈正相关（P＜0.05）;MIC-1、VEGF和P53表达与直肠癌肿瘤临床分期、淋巴结转移呈明显相关性（P＜0.01）;MIC-1和VEGF、P53阳性组和阴性组的5年生存率差异均有显著性（P＜0.01）.结论：MIC-1、VEGF和P53与直肠癌淋巴结转移、分期及预后有密切关系,淋巴结转移是最重要的独立的预后因素.     

Macrophage inhibitory cytokine-1 H6D polymorphism, prostate cancer risk, and survival.

Macrophage inhibitory cytokine-1 (MIC-1), a member of the transforming growth factor-beta superfamily, is important in regulating inflammation. Inflammation of the prostate has been suggested to favor tumor development. A recent study (JNCI 2004, 96:1248-1254) found marginal evidence of an association between the presence of the mature MIC-1 protein nonsynonymous polymorphism H6D C-to-G (rs1058587) with reduced prostate cancer risk [odds ratio, 0.83; 95% confidence interval (95% CI), 0.69-0.99]. We tested this in a population-based study of 819 cases and 731 controls from Australia and found a similar, yet not significant, odds ratio of 0.85 (95% CI, 0.7-1.04; P = 0.11). We also tested the potential association between the H6D variant and disease-specific survival in 640 cases followed-up for an average of 8.2 years. We found that cases carrying the H6D G allele had an increased risk of death from prostate cancer than cases carrying two copies of the C allele (hazard ratio, 1.72; 95% CI, 1.06-2.78; P = 0.03). Our data suggest that the H6D variant in MIC-1 might play a role in prostate cancer, but it is difficult to explain how a variant can be associated with lower risk of developing prostate cancer but more aggressive growth if cancer develops.     

Macrophage inhibitory cytokine-1 induces the invasiveness of gastric cancer cells by up-regulating the urokinase-type plasminogen activator system

In our search for genes associated with gastric cancer progression, we identified macrophage inhibitory cytokine-1 (MIC-1), a member of the transforming growth factor beta superfamily, as an overexpressed gene in gastric tumor tissues. Expression analysis of MIC-1 in gastric tumor tissues revealed a specific expression in gastric cancer cells, and this expression level was well correlated with invasive potential in various human gastric cancer cell lines. Stable transfection of MIC-1 into SNU-216, a human gastric cancer cell line, significantly increased its invasiveness. The overexpression of MIC-1 into SNU-216 cells significantly increased the activity of urokinase-type plasminogen activator (uPA), and the expressions of uPA and urokinase-type plasminogen activator receptor (uPAR). Similarly, the stimulation of gastric cancer cell lines with purified recombinant MIC-1 dose-dependently increased cell invasiveness, uPA activity, and uPA and uPAR expression. However, MIC-1 did not significantly suppress the proliferation of gastric cancer cell lines. We also found that the stimulation of human gastric cell lines with recombinant MIC-1 strongly induced activation of mitogen-activated protein kinase kinase-1/2 and extracellular signal-regulated kinase-1/2. Additional analysis revealed that PD98059, a selective inhibitor of mitogen-activated protein kinase kinase-1/2, suppressed not only gastric cancer cell invasiveness and uPA activity, but also the mRNA expressions of uPA and uPAR, as induced by recombinant MIC-1. Our results indicate that MIC-1 may contribute to the malignant progression of gastric cancer cells by inducing tumor cell invasion through the up-regulation of the uPA activation system via extracellular signal-regulated kinase-1/2-dependent pathway.     

肺癌患者血清中巨噬细胞抑制因子-1水平初探

目的 初步研究肺癌患者血清中巨噬细胞抑制因子-1(MIC-1)的浓度在肺癌临床中的应用价值.方法 采用双抗体夹心ELISA法检测79例肺癌患者、8例肺良性疾病患者及200例正常对照人群血清MIC-1浓度,采用电化学发光免疫分析仪检测上述肺癌患者血清标本CEA、CA125、NSE、SCC、Cyfer21浓度.结果 肺癌组...     

Expression analysis of imbalanced genes in prostate carcinoma using tissue microarrays

To identify candidate genes relevant for prostate tumour prognosis and progression, we performed an exhaustive gene search in seven previously described genomic-profiling studies of 161 prostate tumours, and four expression profiling studies of 61 tumours. From the resulting list of candidate genes, six were selected for protein-expression analysis based on the availability of antibodies applicable to paraffinised tissue: fatty acid synthase (FASN), MYC, beta-adrenergic receptor kinase 1 (BARK1, GRK2) the catalytic subunits of protein phosphatases PP1alpha (PPP1CA) and PP2A (PPP2CB) and metastasis suppressor NM23-H1. These candidates were analysed by immunohistochemistry (IHC) on a tissue microarray containing 651 cores of primary prostate cancer samples and benign prostatic hyperplasias (BPH) from 175 patients. In univariate analysis, expression of PP1alpha (P=0.001) was found to strongly correlate with Gleason score. MYC immunostaining negatively correlated with both pT-stage and Gleason score (P<0.001 each) in univariate as well as in multivariate analysis. Furthermore, a subgroup of patients with high Gleason scores was characterised by a complete loss of BARK1 protein (P=0.023). In conclusion, our study revealed novel molecular markers of potential diagnostic and therapeutic relevance for prostate carcinoma.     

Serum macrophage inhibitory cytokine-1 concentrations correlate with the presence of prostate cancer bone metastases.

Macrophage-inhibitory cytokine-1 (MIC-1) is a divergent member of the transforming growth factor beta superfamily. It is up-regulated by nonsteroidal anti-inflammatory drugs and is highly expressed in human prostate cancer leading to high serum MIC-1 concentrations with advanced disease. A role for MIC-1 has been implicated in the process of early bone formation, suggesting that it may also mediate sclerosis at the site of prostate cancer bone metastases. Consequently, the aim of this study was to retrospectively determine the relationship of serum MIC-1 concentration and other markers related to current and future prostate cancer bone metastasis in a cohort of 159 patients with prostate cancer. Serum markers included cross-linked carboxy-terminal telopeptide of type I collagen, prostate-specific antigen, and amino-terminal propeptide of type I procollagen (PINP). The mean values of all the biomarkers studied were significantly higher in patients with baseline bone metastases (BM+, n = 35), when compared with those without bone metastases (BM-, n = 124). In a multivariate logistic model, both MIC-1 and PINP independently predicted the presence of baseline bone metastasis. Based on receiver operator curve analysis, the best predictor for the presence of baseline bone metastasis was MIC-1, which was significantly better than carboxy-terminal telopeptide of type I collagen, prostate-specific antigen, and PINP. Patients who experienced bone relapse had significantly higher levels of baseline MIC-1 compared with patients who did not (1476.7 versus 988.4; P = 0.03). Current use of acetylsalicylic acid did not influence serum MIC-1 levels in this cohort. Although requiring validation prospectively, these results suggest that serum MIC-1 determination may be a valuable tool for the diagnosis of current and future bone metastases in patients with prostate cancer.     

血清MIC-1和CA125在卵巢上皮癌诊断及预后中的应用研究

目的：探讨巨噬细胞抑制因子-1（MIC-1）和CA125的卵巢癌诊断和预后判断价值。方法通过检测92例未经治疗的卵巢上皮癌患者（观察组）和96例健康女性（对照组）血清样本中的MIC-1和CA125水平,分析血清MIC-1和CA125水平与卵巢癌的关系;通过ROC曲线评价MIC-1和CA125单项及联合应用的卵巢癌诊断效能;通过对其中74例卵巢癌患者按术后残瘤大小和首次化疗敏感性结果分组,比较两组治疗前MIC-1和CA125水平,评价MIC-1和CA125的疗效评价价值;并通过分析68例患者的MIC-1和CA125水平与无瘤生存时间（relapse free survival,RFS）的关系评价其预后判断价值。结果观察组血清MIC-1水平显著高于对照组（P＜0.001）;MIC-1诊断卵巢癌的ROC（AUC=0.945）与CA125的ROC（AUC=0.966）比较差异无统计学意义,MIC-1和CA125联合应用的卵巢癌诊断效能显著提高（AUC=0.966）。术前CA125水平与术后残瘤大小显著相关（P=0.07）;术前血清MIC-1水平与化疗敏感性显著相关（P=0.001）,并且血清MIC-1水平与患者RFS呈负相关。结论 MIC-1和CA125联合检测可提高卵巢癌的诊断率,MIC-1高水平预示卵巢上皮癌患者存在较大的耐药风险,且MIC-1可作为预测卵巢癌患者生存期的独立指标。     

丝氨酸/精氨酸富有剪接因子1、核因子κB在前列腺癌中的表达及与患者临床特征的关系

目的探讨前列腺癌组织中丝氨酸/精氨酸富有剪接因子1(SRSF1)、核因子κB(NF-κB)的表达情况及与患者临床特征的关系。方法选取前列腺癌患者和前列腺良性增生患者,各70例,取前列腺癌组织和前列腺良性增生组织。免疫组化法检测前列腺癌组织和前列腺良性增生组织SRSF1、NF-κB蛋白的阳性表达率,并分析不同临床特征前列腺癌患者前列腺癌组织中SRSF1、NF-κB蛋白的表达情况。结果前列腺癌组织中SRSF1、NF-κB蛋白的阳性表达率均高于前列腺良性增生组织,差异均有统计学意义(P﹤0.05)。不同前列腺特异性抗原(PSA)水平前列腺癌患者前列腺癌组织中SRSF1蛋白的阳性表达率比较,差异无统计学意义(P﹥0.05);不同TNM分期、淋巴结转移情况、Gleason评分前列腺癌患者前列腺癌组织中SRSF1蛋白的阳性表达率比较,差异均有统计学意义(P﹤0.05)。不同PSA水平、Gleason评分前列腺癌患者前列腺癌组织中NF-κB蛋白的阳性表达率比较,差异均无统计学意义(P﹥0.05);不同TNM分期、淋巴结转移情况前列腺癌患者前列腺癌组织中NF-κB蛋白的阳性表达率比较,差异均有统计学意义(P﹤0.05)。结论前列腺癌组织中的SRSF1、NF-κB蛋白表达水平和阳性表达率均较高,SRSF1蛋白的表达可能与TNM分期、淋巴结转移情况和Gleason评分,NF-κB蛋白的表达可能与TNM分期和淋巴结转移情况有关。     

MIC-1/GDF15 in Barrett's oesophagus and oesophageal adenocarcinoma

Background:

Biomarkers are needed to improve current diagnosis and surveillance strategies for patients with Barrett''s oesophagus (BO) and oesophageal adenocarcinoma (OAC). Macrophage inhibitory cytokine 1/growth differentiation factor 15 (MIC-1/GDF15) tissue and plasma levels have been shown to predict disease progression in other cancer types and was therefore evaluated in BO/OAC.

Methods:

One hundred thirty-eight patients were studied: 45 normal oesophagus (NE), 37 BO, 16 BO with low-grade dysplasia (LGD) and 40 OAC.

Results:

Median tissue expression of MIC-1/GDF15 mRNA was ⩾25-fold higher in BO and LGD compared to NE (P<0.001); two-fold higher in OAC vs BO (P=0.039); and 47-fold higher in OAC vs NE (P<0.001). Relative MIC-1/GDF15 tissue expression >720 discriminated between the presence of either OAC or LGD vs NE with 94% sensitivity and 71% specificity (ROC AUC 0.86, 95% CI 0.73–0.96; P<0.001). Macrophage inhibitory cytokine 1/growth differentiation factor 15 plasma values were also elevated in patients with OAC vs NE (P<0.001) or BO (P=0.015).High MIC-1/GDF15 plasma levels (⩾1140 pg ml⁻¹) were an independent predictor of poor survival for patients with OAC (HR 3.87, 95% CI 1.01–14.75; P=0.047).

Conclusions:

Plasma and tissue levels of MIC-1/GDF15 are significantly elevated in patients with BO, LGD and OAC. Plasma MIC-1/GDF15 may have value in diagnosis and monitoring of Barrett''s disease.     

巨噬细胞抑制因子-1及其联合多种标志物在肺癌早期诊断和诊断中的临床意义

目的：研究巨噬细胞抑制因子-1（macrophage inhibitory cytokine-1,MIC-1）在早期肺癌诊断中的辅助价值,并评价多种肿瘤标志物联合应用的临床意义。方法应用MIC-1定量检测试剂盒及Roche Cobas 601电化学发光免疫分析仪分别检测663例未经治疗的不同临床分期的肺癌患者和488例正常人群血清样本中的MIC-1、CEA、CA125、NSE、SCC和CYFRA21-1水平和分布,分析患者血清MIC-1水平与肺癌临床分期、病理分型和细胞分化程度的关系,并研究多种标志物联合检测的价值。结果肺癌患者血清MIC-1水平显著高于正常人群（P＜0.001）;MIC-1水平随临床分期的进展呈上升的趋势（P＜0.001）,且与肿瘤浸润（P＜0.001）、淋巴结转移（P=0.02）、远端转移（P＜0.001）和肿瘤分化程度（P＜0.001）显著相关。单一检测MIC-1诊断肺癌的敏感度（76.6%）高于其他五种肺癌标志物的联合应用（72.2%）,且MIC-1在鳞癌、腺癌、小细胞癌诊断中的敏感度均能达到甚至超过其他五种标志物的联合诊断水平（81.6%vs 82.8%;74.7%vs 68.9%;84.9%vs 83.0%）。在肺癌早期,以MIC-1为主的六种肿瘤标志物联合检测的诊断敏感度（Ⅰ期：79.8%;Ⅱ期：87.7%）显著高于其他五种肿瘤标志物联合诊断的敏感度（Ⅰ期：44.9%;Ⅱ期：72.6%）。结论 MIC-1是肺癌,尤其是早期肺癌有价值的血清肿瘤标志物,MIC-1和CEA、CA125、NSE、SCC、CYFRA21-1联合检测用于普通人群体检和肺癌早期诊断具有重要的临床意义和价值。     

EBAG9/RCAS1 expression and its prognostic significance in prostatic cancer

Estrogen receptor-binding fragment-associated gene 9 (EBAG9) has been identified as a primary estrogen-responsive gene from MCF-7 human breast cancer cells (Watanabe T, et al., Mol Cell Biol 1998;18:442-9). EBAG9 is identical with RCAS1 (receptor-binding cancer antigen expressed on SiSo cells), which has been reported as a cancer cell surface antigen implicated in immune escape (Nakashima M, et al., Nat Med 1999;5:938-42). In our present study, we examined EBAG9 expression in human prostatic tissues and investigated its prognostic significance in patients with prostatic cancer. EBAG9 expression in normal prostatic epithelial cells and PC-3, DU145 and LNCaP cancer cells was determined by Western blot analysis. Immunohistochemic analysis was performed in 21 benign and 81 malignant prostatic specimens, and patients' charts were reviewed for clinical, pathologic and survival data. EBAG9 was abundantly expressed in the prostate cancer cells compared to the normal epithelial cells. Strong and diffuse immunostaining in the cytoplasm of EBAG9 was found in 44 of 81 (54%) cancerous tissue samples. EBAG9 expression significantly correlated with advanced pathologic stages and high Gleason score (p = 0.0305 and < 0.0001, respectively). EBAG9 was more frequently expressed at sites of capsular penetration (79%) and lymph node metastasis (100%) compared to intracapsular primary tumors (54%) (p = 0.0264 and 0.0048, respectively). Positive EBAG9 immunoreactivity significantly correlated with poor PSA failure-free survival (p = 0.0059). EBAG9/RCAS1 may play a significant role in cancer progression via an immune escape system. Immunodetection of EBAG9/RCAS1 expression can be a negative prognostic indicator for patients with prostatic cancer.     

Expression of prostate specific membrane antigen and three alternatively spliced variants of PSMA in prostate cancer patients

Prostate specific membrane antigen (PSMA) is a folate gamma glutamyl carboxypeptidase that is oriented on the plasma membrane of normal and prostate cancer cells. A cytosolic version of PSMA, PSM', results from alternative splicing of the PSMA gene. Two additional alternatively spliced variants of PSMA, PSM-C and PSM-D, have been described recently. The ratio of PSMA to PSM' mRNA was higher in a small number of prostate cancer specimens compared to normal prostate cancer and benign prostatic hypertrophy (Su et al. Cancer Res 1995;55:1441). The intent of our study was to measure the gene expression of PSMA and the 3 PSMA splice variants in a large number of patient's tissues. A real-time, quantitative PCR assay was developed to quantify PSMA, PSM', PSM-C and PSM-D. Discrimination among the variants was achieved by designing unique primers and TaqMan probes for each gene. Amplification and detection was specific for the desired splice variant and was sensitive to one gene copy per reaction. The assay was used to quantify the gene expression in specimens of normal, benign, primary and metastatic prostate cancer from 72 patients. The mean PSMA expression (relative to 18S rRNA) was 2- to 3-fold lower in normal prostate (n = 4) compared to primary (n = 55, p = 0.31) and metastatic (n = 20, p = 0.33) prostate cancer. There was no difference in the PSMA expression between benign and cancerous prostate tissue from the same patients (n = 35). The ratio of PSMA to PSM' was lowest in the normal prostate and increased with increasing Gleason score (p < 0.001). The increased ratio in these tissues was a reflection of both increasing PSMA levels and decreasing PSM' mRNA. The expression of PSM-C did not differ in any of the tissue categories studied. The expression of PSM-D was similar in normal and primary prostate cancer but was 2-fold higher in lymph node (p < 0.005) and bone metastases (p < 0.05) compared to the primary tumors. Our results of the first detailed quantitative analysis of PSMA mRNA expression in patient's tissues demonstrate that PSMA and the 3 PSMA splice variants are expressed in normal, benign, cancerous and metastatic prostate cancer. We note increased PSMA expression in some malignant tissues, however, these increases are modest in magnitude. We also report that the expression of a novel splice variant, PSM-D, is elevated in prostate cancer metastases.     

长链非编码RNA TUG1在前列腺癌组织中的表达及与预后的关系

目的:探究长链非编码核糖核酸（lncRNA）牛磺酸上调基因1（TUG1）在前列腺癌（PCa）患者癌组织中的表达水平及与预后的关系。方法:以本院收治的54例PCa患者为研究对象,取癌组织和癌旁正常组织,采用qRT-PCR检测lncRNA TUG1表达水平,分析TUG1表达与PCa患者临床病理参数的关系,Kaplan-Meier分析PCa患者生存情况,COX回归分析影响PCa患者预后的危险因素。结果:TUG1在PCa患者癌组织中的表达水平显著高于癌旁正常组织（P＜0.05）;PCa患者癌组织中TUG1表达水平与患者年龄、淋巴结转移情况、T分期、前列腺特异性抗原（PSA）水平和Gleason评分有关（P＜0.05）;Kaplan-Meier法分析结果显示TUG1低表达组PFS、OS均显著高于高表达组（P＜0.05）;COX分析结果显示TUG1表达、PSA水平和Gleason评分是影响PCa患者预后的独立危险因素。结论:lncRNA TUG1在PCa患者癌组织中高表达,与PCa发生及发展有关,可作为临床评估患者预后的参考指标。     

Role of macrophage inhibitory cytokine-1 in tumorigenesis and diagnosis of cancer

Macrophage inhibitory cytokine-1 (MIC-1), a transforming growth factor-beta superfamily cytokine, is involved in tumor pathogenesis, and its measurement can be used as a clinical tool for the diagnosis and management of a wide range of cancers. Although generally considered to be part of the cell's antitumorigenic repertoire, MIC-1 secretion, processing, and latent storage suggest a complex, dynamic variability in MIC-1 bioavailability in the tumor microenvironment, potentially modulating tumor progression and invasiveness.     

细胞分化抑制子Id1在前列腺癌中的表达及意义

目的:细胞分化抑制子Id1(inhibitor of differentiation or inhibitor of DNA binding)蛋白与肿瘤的发生、侵袭及远处转移有关。本研究主要观测人前列腺癌组织中Id1蛋白的表达,并分析其应用价值。方法:采用免疫组化SP法检测43例前列腺癌(prostate cancer,PC)、12例前列腺增生(benign prostatic hyperplasia,BPH)和2例正常前列腺组织中Id1蛋白的表达。结果:Id1蛋白在正常前列腺组织中无表达;BPH组织中无或弱表达(5/12);所有前列腺癌组织都有阳性表达(43/43),而且大多数以中、高度表达为主(与BPH相比,P<0.01);前列腺癌中Id1的表达水平与Gleason分级成正比(rs=0.63P<0.01)。结论:人前列腺癌组织中,Id1蛋白有过度表达,且与前列腺癌的Gleason分级呈正相关,表明与癌组织的恶性程度有关。Id1蛋白有可能作为判断前列腺癌发展的一种新的肿瘤标志。