Expression of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) as a prognostic indicator in gastric cancer

In this study the expression levels of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) in gastric cancer cell lines and tissues have been analysed in order to assess their value as a prognostic indicator. The expressions of RECK, activated matrix metalloproteinase (MMP)-7, and vascular endothelial growth factor (VEGF) in gastric cancer tissues and cell lines were evaluated by Western blot analysis; and MMP-2 and MMP-9 were evaluated by gelatin zymography. RECK expression in the context of gastric cancer was also compared with various clinicopathologic parameters and compared to the expression of activated MMP-7, MMP-2, and MMP-9. Fifty-two percent of the 102 gastric cancer tissues and 81.8% of the 11 gastric cancer cell lines exhibited reduced RECK expression. We also detected a significant inverse correlation between RECK expression and macroscopic tumour growth (P=0.018), lymphatic invasion (P=0.018), lymph node metastasis (P=0.000), stage (P=0.000), and MMP-9 (P=0.039). No correlation between RECK expression and MMP-7 and MMP-2, VEGF were detected. Our data strongly supports the hypothesis that RECK is a suppressor of malignancy, and constitutes a good prognostic indicator in gastric cancer.     

CD147, MMP-2, MMP-9 and MVD-CD34 are significant predictors of recurrence after liver transplantation in hepatocellular carcinoma patients

OBJECTIVE: The prognostic role of tumor biological markers (biomarkers) in predicting recurrence of hepatocellular carcinoma (HCC) was investigated in this study, the results of which may help to select appropriate candidates for liver transplantation (LT). METHODS: Tissue samples from 82 HCC patients with cirrhosis who had undergone LT were immunohistochemically stained with antibodies of anti-CD147, anti-matrix metalloproteinases-2 (MMP-2), MMP-9 and anti-vascular endothelial growth factor (VEGF). Tumor microvessel density (MVD) was evaluated by using CD34. Multivariate Cox regression analysis was performed to identify the relevant prognostic factors. RESULTS: A significant correlation was found between the expression of CD147, VEGF, MMP-2, MMP-9 and MVD-CD34 in HCC. Tumor CD147 expression (P < 0.0001), tumor MVD-CD34 (P < 0.0001), MMP-9 in stromal compartment (P = 0.0257) and tumor VEGF expression (P = 0.0335) were significantly associated with the recurrence in HCC patients after LT. Univariate analysis showed that strong CD147 expression and high MVD-CD34 were significantly associated with poor tumor recurrence-free survival after LT (P < 0.0001). Multivariate analysis indicated that CD147 (P = 0.0001), MVD-CD34 (P = 0.0118), MMP-2 (P = 0.0312) and MMP-9 (P = 0.0280) in stromal compartment were all significant predictors in predicting HCC recurrence, while VEGF, MMP-2 and MMP-9 in tumor compartment were not significantly associated with poor prognosis. CONCLUSIONS: The tumor biomarkers CD147 and MVD-CD34 are more feasible markers for rational selection of LT candidates with HCC. MMP-9 and MMP-2 expression in stromal compartment, combined with pTNM tumor grade, may be helpful in predicting poor prognosis in HCC patients after LT.     

Correlation of reversion-inducing cysteine-rich protein with kazal motifs (RECK) and extracellular matrix metalloproteinase inducer (EMMPRIN), with MMP-2, MMP-9, and survival in colorectal cancer

mRNA, and latent and active levels MMP-2 and -9 were higher in tumor tissue compared to normal tissue from 63 patients with colorectal cancer, whereas RECK and EMMPRIN levels were lower. Correlations between mRNA, latent, and active MMP were particular high for MMP-2 in tumor tissue (R(s)=0.6-0.8, P<0.001). For active MMP-2, but not for MMP-9, a significant negative partial correlation (R(p)=-0.440, P<0.001) for RECK was found in tumor tissue, which was confirmed by linear regression analysis. In exploratory survival analyses we found that in patients with localized disease the RECK level in normal or tumor tissue had a significant (P=0.017) association with overall survival.     

Expression of a novel matrix metalloproteinase regulator, RECK, and its clinical significance in resected non-small cell lung cancer

The reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) was initially isolated as a transformation-suppressor gene by expression cloning and found to encode a membrane-anchored regulator of the matrix metalloproteinases (MMPs). Experimental studies have shown that RECK can suppress tumour - invasion, metastasis and angiogenesis. However, the clinical impact of RECK remains unclear. To assess the clinical significance of RECK-expression in non-small cell lung cancer (NSCLC), a total of 171 patients with completely resected pathological stage (p-stage) I-IIIA NSCLC were retrospectively examined. Expression of RECK and vascular endothelial growth factor (VEGF) in tumour tissues was assessed by immunohistochemical staining (IHS). Intratumoural microvessel density (IMVD), a measurement of angiogenesis, was also determined by IHS using an anti-CD34 antibody. A significant inverse correlation between RECK-expression and tumour angiogenesis was documented; the mean IMVD in tumours with strong RECK-expression (157.1) was significantly lower than that observed in tumours with weak RECK-expression (194.5; P = 0.008). Interestingly, this inverse correlation was seen only when VEGF was strongly expressed, which suggests that RECK could suppress the angiogenesis induced by VEGF. The 5-year survival rate for patients with tumours with strong RECK-expression (75.8%) was significantly higher than that for patients with weakly expressing tumours (54.3%; P = 0.016). Subset analyses showed that the prognostic impact of RECK-status was evident in patients with either adenocarcinoma, poorly differentiated tumours, or p-stage IIIA disease. A multivariate analysis confirmed that reduced RECK-expression was an independent and significant factor in predicting a poor prognosis (P = 0.009; Hazard ratio (HR), 0.474 with a 95% Confidence interval (CI) of 0.271-0.830). In conclusion, RECK-status is a significant prognostic factor correlated with tumour angiogenesis in NSCLC patients.     

TACE联合血管内皮抑制素治疗肝癌的短期疗效及其对患者血清VEGF、MMP-9、OPN的影响

目的:探讨经肝动脉化疗栓塞（TACE）联合血管内皮抑制素治疗肝癌的短期疗效及其对患者血清VEGF、MMP-9、OPN的影响。方法:选取2012年4月-2013年4月我院收治的原发性肝细胞癌（简称肝癌）患者80例,按照随机对照法分为观察组和对照组各40例,其中观察组给予TACE联合重组人血管内皮抑制素（恩度）治疗,对照组采用单纯的TACE治疗,观察两组患者治疗前、治疗后1、3、7、15、30d血清血管内皮生长因子（VEGF）、基质金属蛋白酶9（MMP-9）、骨桥蛋白（OPN）水平的变化,观察两组患者肿瘤新生血管抑制情况、肿瘤控制情况及1年生存率。结果:治疗前两组的血清VEGF、MMP-9、OPN水平比较差异无统计学意义（P＞0.05）;治疗后各时间节点观察组血清VEGF、MMP-9、OPN水平与治疗前比较差异无统计学意义（P＞0.05）;而对照组治疗后各时间节点血清VEGF、MMP-9、OPN水平均较治疗前明显升高,差异具有统计学意义（P＜0.05）;观察组患者新生血管控制率、疾病控制率（DCR）及术后1年生存率均明显优于对照组,差异具有统计学意义（P＜0.05）。两组的不良反应发生率比较差异无统计学意义（P＞0.05）。结论:TACE联合血管内皮抑制素治疗肝癌可以有效抑制肿瘤新生血管的形成,降低肿瘤复发转移的几率,延长患者的生存时间,提高生存率,其作用机制可能与抑制TACE术后血浆VEGF、MMP-9、OPN水平有关。     

RECK—a newly discovered inhibitor of metastasis with prognostic significance in multiple forms of cancer

The RECK (reversion-inducing cysteine rich protein with Kazal motifs) protein was initially discovered by its ability to induce reversion in ras-activated fibroblasts. The key action of RECK is to inhibit matrix metalloproteinases (MMPs) involved in breakdown of the extracellular matrix (ECM), and angiogenesis—namely MMP-2, MMP-9 and MTP-1. To this effect, it plays important physiological roles in embryogenesis and vasculogenesis. Additionally, it has a significant effect on tumorigenesis by limiting angiogenesis and invasion of tumours through the ECM. RECK has been studied in the context of a number of human tumours including colorectal, breast, pancreas, gastric, hepatocellular, prostate, and non-small cell lung carcinoma. In many of these tumours, RECK is down-regulated most likely as a result of inhibition at the Sp1 promoter site. MMP-2 and MMP-9 generally show an inverse association with RECK expression, but there are exceptions to this rule. Likewise, a reduction in tumour microvascular density (MVD) and VEGF have also been correlated with increased RECK levels, although more studies are required to define this effect. The predominant finding across all human tumour studies is a significantly improved prognosis (due to decreased invasion and metastasis) in tumours with preserved RECK expression. Although further research is required, RECK is a promising prognostic marker and potential therapeutic agent in multiple cancers.     

塞来昔布对实验性结肠癌原位移植瘤生长及血管形成的影响

目的:探讨塞来昔布对实验性结肠癌原位移植瘤生长及血管形成的影响。方法:使用对数生长期的人结肠癌细胞(HT-29)于裸鼠皮下接种成瘤后原位种植。术后随机分为对照组(C组)及塞来昔布高、中、低剂量组(H、M、L组)共四组进行研究。结果:24只裸鼠实验期间无1只死亡,成瘤率为100%,比较各组原位移植瘤体积和瘤质量差异有统计学意义,P值均<0·05。L、M和H组的抑瘤率分别为25·30%、38·80%和76·92%,与对照组比较差异有统计学意义,P=0·000,且存在明显的剂量依赖。干预组瘤组织中MVD、VEGFmRNA、MMP-2mRNA和匀浆上清中PGE2含量与对照组相比差异有统计学意义,P值分别为0·050、0·050、0·050和0·010,随着剂量增加,MVD、VEGFmRNA、MMP-2mRNA和匀浆上清中PGE2含量逐渐降低。瘤组织中PGE2含量与瘤质量,以及MVD与VEGFmRNA、MMP-2mRNA均存在显著的正相关性,r=0·8814,P=0·000;r=0·8573,P=0·000;r=0·6427,P=0·001。结论:塞来昔布通过抑制人结肠癌裸鼠原位移植瘤环氧化酶-2活性,抑制PGE2合成、VEGFmRNA和MMP-2mRNA表达,抑制肿瘤的血管形成,从而对结肠癌的生长有抑制作用。     

脑膜瘤中MMP-9、VEGF表达及其与肿瘤血管新生及复发的关系

目的：探讨脑膜瘤组织中基质金属蛋白酶-9（matrix metallo proteinases-9,MMP-9）、血管内皮生长因子（vascular endothelial growth factor,VEGF）的表达及其与微血管密度（microvessel density,MVD）及肿瘤复发的关系.方法：采用SP法,分别检测78例脑膜瘤病理标本中VEGF,MMP-9表达及MVD情况,并分析在脑膜瘤中VEGF,MMP-9表达及其与MVD、肿瘤病理分级、复发情况之间的关系.结果：脑膜瘤中MMP-9、VEGF的阳性表达率分别为62.82％、75.64％,脑膜瘤中MMP-9与VEGF表达具有正相关关系（r=0.449,P＜0.01）;脑膜瘤中MMP-9、VEGF的表达与MVD值呈显著正相关（r=0.692,P=0.000;r=0.755,P=0.000）;脑膜瘤中MMP-9、VEGF表达及MVD与肿瘤病理分级、复发有显著相关性（P＜0.01）.结论：脑膜瘤组织中MMP-9、VEGF蛋白的表达与肿瘤组织的血管生成、恶性程度及复发情况密切相关,可以作为肿瘤恶性程度及复发潜能的参考指标.     

AEG-1、MMP-9在结直肠组织中的表达及临床病理相关性的研究

目的 探讨星形细胞上调基因1(AEG-1)和基质金属蛋白酶9(MMP-9)在结直肠组织中的表达并分析其与结直肠癌的临床病理因素之间的相关性。方法 应用免疫组化技术检测AEG-1和MMP-9在结直肠正常黏膜(45例)、低级别腺瘤(31例)、高级别腺瘤(15例)和结直肠癌(146例)中的表达,并分析其表达的意义。结果 AEG-1在结直肠正常黏膜、低级别腺瘤、高级别腺瘤和结直肠癌中有逐步升高的趋势。MMP-9表达与AEG-1有类似的趋势。AEG-1在结直肠癌组织中的阳性表达率与临床分期、T分期、N分期、M分期及组织分化有统计学差异(P＜0.05),而与年龄、性别、肿瘤位置和肿瘤大小无统计学差异(P＞0.05)。在结直肠癌中,AEG-1和MMP-9表达呈显著正相关,AEG-1、MMP-9高表达的患者具有较短的生存期(P＜0.05)。结论 AEG-1参与结直肠癌的发生,与结直肠癌的演进相关,并可能作为结直肠癌患者预后的一种新的生物标志物。     

结直肠肿瘤微血管计数及血管内皮细胞生长因子表达

目的 探讨血管生成与结直肠肿瘤的发生,发展关系,评估微血管计数（ＭＶＤ值）及血管内皮细胞生长因子（ＶＥＧＦ）表达与结直肠肿瘤预后的相关性。方法 应用免疫组化法回顾性地研究了３２例结直肠肿瘤石蜡包埋的病理组织。结果 正常粘膜,腺瘤,癌组织的ＭＶＤ值递增。不同病理状态下的结直肠癌ＭＶＤ值有差异,ＶＥＧＦ阴性组ＭＶＤ值低于ＶＥＧＦ阳性组,低ＭＶＤ值主ＶＥＧＦ阴性组生存率高于高ＭＶＤ值组及ＶＥＧＦ阳性组。     

RECK及MMP-14在肝细胞肝癌组织中的表达及其临床意义

目的：探讨RECK及MMP-14与肝细胞肝癌（HCC）的侵袭转移以及预后的关系。方法：用逆转录聚合酶链反应（RT-PCR）技术检测61例HCC患者癌组织、癌旁肝组织以及18例正常肝组织中RECK及MMP-14的mR-NA表达情况;并分析RECK及MMP-14的mRNA的表达与相关临床参数的关系。结果：RECKmRNA在HCC组织中的表达水平显著低于在癌旁肝组织和正常肝组织中的表达水平（P〈0.01）,而在癌旁肝组织和正常肝组织中的表达水平差异无显著性（P〉0.05）;MMP-14mRNA在肝癌组织的表达水平显著高于在癌旁肝组织中的表达水平（P〈0.01）,在癌旁肝组织中的表达水平显著高于在正常肝组织中的表达水平（P〈0.01）。RECKmRNA和MMP-14mRNA在HCC组织中的表达呈负相关（P=0.027）,而两者在癌旁肝组织及正常肝组织中的表达无相关性。RECKmRNA在HCC组织中的表达水平与临床分期、门静脉癌栓、肝外转移及术后复发等明显相关,而与肿瘤直径、肿瘤个数、血清AFP水平、肿瘤分化程度以及癌旁有无肝硬化等无明显关系。MMP-14mRNA在HCC组织中的表达水平与临床分期、门静脉癌栓、肝外转移、术后复发、肿瘤直径大小明显相关,而与肿瘤数目、血清AFP水平、肿瘤分化程度以及癌旁有无肝硬化等无明显关系。结论：本研究提示RECK及MMP-14的mRNA表达与HCC的浸润转移有关,RECK及MMP-14可能在肝细胞肝癌的发生、发展中起重要作用,有可能作为预测肝癌复发、转移的参考指标。     

VEGF与MMP-9在肝癌组织中的表达及其临床意义

背景与目的：血管内皮生长因子vascular endothelial growth factor,VEGF）及基质金属蛋白酶-9（matrix metalloproteinase-9,MMP-9）与恶性肿瘤的侵袭、血管生成及预后有一定关系。VEGF与MMP-9表达对预测肝细胞癌（以下简称“肝癌”）患者预后的报道结果不一。本研究旨在探讨VEGF与MMP-9在肝癌组织中的表达及其预测肝癌患者预后的意义。方法：按是否复发将80例肝癌患者分为复发组与未复发组,采用免疫组化方法检测两组手术切除标本中VEGF及MMP-9的表达情况,分析两者的表达与临床病理指标的关系。建立Cox比例风险模型进行肝癌术后复发风险的多因素分析。结果：MMP-9及VEGF表达于肿瘤细胞浆中,复发组的阳性率分别为50．0％（24／48）及87．5％（42／48）,未复发组分别为15．6％（5／32）及59、4％（19／32）,差异均有显著性（P〈0．05）。VEGF与MMP-9的表达呈正相关（rs=0．36,P〈0．01）,并均与复发呈正相关（P〈0．01）。VEGF阴性患者的1、2、3年累积无瘤生存率分别是85．7％、71．4％及66．3％,VEGF阳性患者分别为58．0％、38．9％及33．9％,差异有显著性（P〈0．01）;MMP-9阴性组分别为72．4％、63．8％及55．5％,MMP-9阳性组分别为50．0％、14．1％及14．1％,差异有显著性（P〈0．01）。多因素分析发现,肝癌组织中VEGF和MMP-9的表达、术前播散结节及镜下微转移灶是肝癌术后复发的独立危险因素。结论：肿瘤组织中VEGF及MMP-9的表达与肝癌患者的术后复发密切相关,为预测肝癌患者术后复发的指标之一。     

晚期非小细胞肺癌患者外周血VEGF和bFGF及MMP-9水平与预后的关系

目的 检测晚期非小细胞肺癌（NSCLC）患者化疗前后外周血血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）和基质金属蛋白酶-9（MMP-9）的水平,探讨其与疗效、预后的关系。方法 酶联免疫吸附试验（ELISA）检测46例晚期NSCLC患者化疗前后血浆VEGF、bFGF和MMP-9水平,并对随访资料进行生存分析,应用COX回归模型分析预后影响因素。结果 化疗前血浆VEGF、bFGF和MMP-9中位值分别为275pg／ml、69pg／ml和122ng／ml,均高于健康对照组（P〈0．05）。化疗前血浆VEGF与bFGF水平正相关,Spearman’S相关系数为0．329。化疗前血浆VEGF、bFGF和MMP-9水平与白细胞、血色素、血小板计数无相关性,与年龄、性别、病理类型、分化程度、TNM分期等亦无关。Ⅳ期广泛转移者（包括骨转移）化疗前血浆MMP-9水平显著高于仅有骨转移者（P=0．013）。化疗后血浆bFGF下降为预后的独立保护因素,RR=11．737（P=0．02）。结论 检测晚期NSCLC患者外周血的某些血管生成相关因子,可能有利于协助预测转移倾向及评价预后。     

Determining the levels of matrix metalloproteinase-9 in portal and peripheral blood is useful for predicting liver metastasis of colorectal cancer

BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is one of the MMPs that play an important role in cancer invasion and metastasis. Increased levels of MMP-9 in tumor tissue have been found to correlate with advanced stages of colorectal cancer. However, the clinical significance of determining the levels of MMP-9 in blood samples from patients with colorectal cancer has not yet been clarified. The purpose of this study was to clarify the relationship between the clinicopathological variables of colorectal cancer and MMP-9 levels of drainage (portal) or peripheral venous blood and to examine whether this assay would be useful for predicting liver metastasis. METHODS: Blood samples were obtained from peripheral and drainage veins of 102 patients with colorectal cancer during surgery and the plasma levels of MMP-9 were determined by a one-step sandwich enzyme immunoassay. RESULTS: The levels of portal MMP-9 were significantly higher than those of peripheral blood (P < 0.01, n = 102). The levels of MMP-9 in peripheral venous blood did not correlate with any of the 12 clinicopathological variables examined, while the levels of MMP-9 in portal blood correlated with macroscopic type of the primary tumor (P = 0.02), Dukes' stage (P = 0.03), liver metastasis (P < 0.01) and lymph node metastasis (P = 0.02). By setting the cutoff ratio of portal to peripheral MMP-9 levels at 1.6 in patients with curative resection (n = 73), elevated ratios predicted subsequent emergence of liver metastases with 77.8% sensitivity, 81.3% specificity and 80.8% accuracy. CONCLUSION: The results suggest that synchronous determination of the levels of MMP-9 in portal and peripheral blood would be useful for selecting colorectal cancer patients at high risk of hepatic recurrence.     

RECK expression in pancreatic cancer: its correlation with lower invasiveness and better prognosis.

BACKGROUND: The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene was initially isolated as a transformation suppressor gene. The RECK gene is expressed widely in normal organs but is undetectable in many tumor-derived cell lines. When artificially expressed in such cell lines, RECK negatively regulates at least matrix metalloprotease (MMP)-9, MMP-2, and MT1-MMP activation and suppresses the invasive and metastatic potentials of these cells. Clinical relevance of these observations, however, is yet to be established. The aim of this study was to examine RECK expression in pancreatic cancer, where intensive invasiveness and metastasis are frequently observed, and investigate its clinical significance. We also analyzed the correlation between RECK expression and MMP activation. METHODS: (a) RECK expression in surgically resected tissue samples of invasive ductal carcinomas of the pancreas (n = 50) was examined immunohistochemically, and its correlation with clinicopathological factors was analyzed; and (b) gelatin zymography was used for the detection of latent and activated forms of MMP-2 and MMP-9 in some of the tissue samples (n = 33). The gelatinase activity was quantified by densitometory, and the ratio of intensity of the active MMP-2 band to the total intensity of the pro- and active MMP-2 bands was evaluated as an indicator of MMP-2 activation. The MMP-9 activation was also studied. RESULTS: Among the 50 ductal carcinoma samples, 26 (52%) were stained positive for RECK. In the normal pancreas, both acinar and beta cells were stained positive, but ductal cells did not. Tumors with positive RECK staining were significantly less invasive as compared with RECK-negative tumors (P = 0.0438). Importantly, patients who had tumors with high RECK expression showed significantly better prognosis than those who had RECK-negative tumors (P = 0.0463, by Log-rank test). Zymographic analysis indicated significant inverse correlation between the level of RECK expression and extent of MMP-2 activation (P = 0.0374). CONCLUSIONS: Our findings support the hypothesis that the RECK protein has negative effects on the invasiveness of pancreatic cancer by inhibiting MMP-2 activation and suggest the potential value of RECK as a prognostic molecular marker for pancreatic cancer.     

Low expression of RECK indicates a shorter survival for patients with invasive breast cancer

Expression cloning was used to initially isolate the reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene as a suppressor of transformation. The gene was found to encode a membrane-anchored regulator of MMPs. Experimental studies showed that RECK can suppress tumor invasion, metastasis, and angiogenesis. However, the clinical impact of RECK remains unclear. To assess the clinical significance of RECK expression in invasive breast cancer, a total of 119 patients with invasive breast cancer were retrospectively examined. Expression of RECK in tumor tissues was assessed by immunohistochemical staining. A significant correlation between RECK expression and 5-year survival rate was documented. The 5-year survival rate for patients with strong RECK expression was significantly higher than that for patients with weakly expressing tumors. Univariate and multivariate analyses confirmed that reduced RECK expression was an independent and significant factor in predicting a poor prognosis. In conclusion, RECK expression is a significant prognostic factor correlated with long-term survival for patients with invasive breast cancer. RECK expression is therefore a potentially useful prognostic marker for breast cancer.