MT1-MMP在肿瘤血管新生中的作用

基质金属蛋白酶 (MMPs)是包含至少 2 6个成员的酶家族。它们除降解细胞外基质外 ,还可以作用于其它的蛋白激酶、细胞趋化分子、生长因子、生长因子连接蛋白以及细胞表面分子等。MMPs的活性受组织型金属蛋白酶抑制物 (TIMPs)调控。MMPs和TIMPs在生理状态和癌症进程中的细胞外基质重塑过程中发挥重要作用 ,特别是MMP 2和TIMP 2。它们主要是调节肿瘤进展的早期阶段 ,如肿瘤生成和血管新生。     

胎膜中MMP-2/TIMP-2表达与胎膜早破关系的研究

目的探讨基质金属蛋白酶-2（MMP-2）及其抑制因子-2（TIMP-2）在自发性胎膜早破孕妇的胎膜中的变化及其意义。方法收集胎膜早破确诊病例30例,同期收集正常妊娠择期剖宫产30例。采用免疫组织化学PV9000法检测MMP-2及TIMP-2在胎膜中的表达。结果1.所选两组标本感染率比较差别无统计学意义（χ^2=2.857,P=0.091）;2.PROM组胎膜组织结构明显疏松,羊膜与绒毛膜之间的区域普遍较对照组增宽;3.早破组与正常组在HE染色后着色深浅有异,早破组略显淡;4.MMP-2在胎膜早破产妇胎膜中的表达高于正常妊娠剖宫产组阳性率分别为100%,46.67%差异有显著性（U=5.849,P=0.000）;5.TIMP-2在胎膜早破产妇胎膜中的表达低于正常妊娠剖宫产组,差异无显著性（U=1.690,P=0.091）。结论胎膜早破的发生可能与MMP-2的表达上调有关。     

胰腺癌MT1-MMP和MMP-2表达与神经浸润、转移及预后关系

目的 探讨膜型基质金属蛋白酶-1(MTI-MMP)和基质金属蛋白酶-2(MMP-2)在胰腺癌中的表达及其与神经浸润、转移及预后的关系.方法 采用免疫组化双标技术检测47例胰腺癌组织中MT1-MMP和MMP-2的表达,并以S-100标记神经纤维,观察MT1-MMP和MMP-2的阳性表达率与胰腺癌神经浸润、转移及预后的关系.结果 MT1-MMP和MMP-2在有神经浸润的胰腺癌组织中阳性表达率均明显高于无神经浸润者(χ2=4.24,11.57;P<0.05);MT1-MMP和MMP-2的表达与胰腺癌转移、临床分期和预后有关(χ2=7.42,7.26;11.85,12.69;6.69,7.86;P<0.01),而与性别、年龄、肿瘤大小及组织学分级无关(χ2=0.05～4.29;P>0.05);胰腺癌组织中MT1-MMP和MMP-2的表达呈正相关(r＝0.585,P<0.01).结论MT1-MMP和MMP-2在胰腺癌的神经浸润、转移过程中可能发挥协同作用;MT1-MMP和MMP-2的表达水平与胰腺癌临床分期和预后密切相关,可作为反应胰腺癌生物学行为的重要指标.     

母血、羊水、脐血中MMP-9及其组织抑制物TIMP-1水平变化与胎膜早破关系的临床研究

目的探讨母血、羊水、脐血中基质金属蛋白酶9(MMP-9)及其组织抑制物1(TIMP-1)的水平变化与胎膜早破发生的相关性,分析其作为一种新型的生物学标志物对绒毛膜羊膜炎和新生儿预后判断的临床价值.方法采用双抗体夹心酶联免疫吸附法检测58例胎膜早破母血、羊水、脐血中MMP-9/TIMP-1水平的水平变化,同时进行胎膜的病理检查.结果胎膜早破组母血、羊水、脐血中MMP-9的含量均高于对照组,尤以羊水中MMP-9水平升高更明显,而TIMP-1水平变化则明显低于对照组(P＜0.05,P＜0.01).MMP-9的含量随破膜时间的延长而增高,尤其破膜时间超过24h增高更为明显.而TIMP-1则随破膜时间的延长而下降,尤其破膜时间超过24h降低更为明显.并发绒毛膜羊膜炎者其MMP-9水平明显高于非绒毛膜羊膜炎患者,而TIMP-1水平则明显低于非绒毛膜羊膜炎患者(P＜0.05,P＜0.01).胎膜早破组产妇所生新生儿Apgar评分≤7分者,其MMP-9的含量显著高于Apgar≥8分的新生儿,而TIMP-1的含量则显著低于Apgar≥8分的新生儿(P＜0.05,P＜0.01).结论MMP-9的异常升高及其抑制物TIMP-1的显著下降是胎膜早破发生的重要发病机制.MMP-9/TIMP-1含量变化可以成为一种新的生物学标志物用于胎膜早破并绒毛膜羊膜炎特别是尚处于亚临床感染状态的孕妇进行早期诊断,并有助于评估新生儿预后.     

MMP-9、IL-6在孕妇羊水中的水平与胎膜早破

目的探讨金属基质蛋白酶-9(MMP-9)、白细胞介素-6(IL-6)在胎膜早破(PROM)孕妇羊水中的改变.方法采用酶联免疫吸附法检测胎膜早破孕妇26例,平均孕期(37.83±0.65)w,平均年龄(27.86±2.30)岁;选取同期住院正常妊娠妇女15例为对照组,平均孕期(38.16±0.76)w,平均年龄(28.06±2.01)岁.结果膜早破孕妇羊水中MMP-9、IL-6水平明显高于对照组,差异显著(P＜0.01、P＜0.01);随着破膜时间的延长羊水中MMP-9、IL-6水平有增高趋势,尤其是破膜时间超过16h者明显增高,差异显著(P＜0.01、P＜0.01).羊水中MMP-9和IL-6水平,在胎膜早破合并绒毛膜羊膜炎与非绒毛膜羊膜炎患者,差异显著(P＜0.05和P＜0.01).结论MMP-9、IL-6可作为一个敏感的新指标,为预防胎膜早破的发生提供了临床依据.     

早期妊娠的丢失与MMP-2、MMP-9及TIMP-1、TIMP-2的研究进展

基质金属蛋白酶（matrix metalloproteinases,MMPs）是参与细胞外基质（extracellular matrix,ECM）降解和重塑正常组织的重要酶类,基质金属蛋白酶的组织抑制物（tissue inhibitor of metalloproteinases,TIMPs）是MMPs活性的特异性抑制剂,在正常情况下MMPs和TIMPs维持着一个动态平衡。     

母血IL-2在胎膜早破中的变化

目的探讨胎膜早破时母血中Th1因子IL-2水平的变化趋势及意义。方法酶联免疫吸附方法检测20例胎膜早破（PROM）其中8例合并重度妊娠高血压疾病、12例无合并症者,20例正常孕妇组并比较两组孕妇血清IL-2含量。结果PROM组IL-2在母血中含量高于对照组（P〈0.05）。PROM无合并症组低于合并妊娠高血压疾病组（P〈0.05）。结论PROM中孕妇血清中IL-2含量升高。     

基质金属蛋白酶MMP-2、MMP-9及其抑制因子TIMP-1、TIMP-2在子宫内膜异位症血清中的表达及临床意义

目的探讨基质金属蛋白酶MMP-2、MMP-9及其抑制因子TIMP-1、TIMP-2在子宫内膜异位症（EMs）血清中的表达及临床意义。方法采用双抗体夹心酶联免疫吸附法（ELISA）检测55例EMs患者和30例对照组血清中删P-2,MMP-9、TIMP-1和TIMP-2水平。结果BMs组血清中MMP-2和MMP-9水平显著高于对照组,TIMP-1和TIMP-2水平显著低于时照组（P〈0．05）。Ⅲ-Ⅳ期血清MMP-2和MMP-9水平显著高于Ⅰ-Ⅱ期和对照组,TIMP-1和TIMP-2水平显著低于Ⅰ-Ⅱ期和对照组（P〈0．05）。结论MMP-2、MMP-9与Elds发生和发展相关,TIMP-1、TIMP-2对MMP-2、MMP-9水平调节有重要作用。     

解脲支原体感染在胎膜早破中的作用

目的探讨解脲支原体感染与胎膜早破的关系.方法采用培养法分别对52例胎膜早破和30例正常妊娠妇女的宫颈分泌物、胎膜组织进行UU检测,并对两组胎盘胎膜组织进行常规病理检查.结果观察组宫颈分泌物和胎膜组织中,UU检出率明显高于对照组,两组比较,差异有显著性(P＜0.05,P＜0.005).观察组胎盘绒毛膜羊膜炎明显高于对照组,差异有显著性(P＜0.005).结论UU感染与胎盘绒毛膜羊膜炎和胎膜早破有关.     

IL-6、MMP-9、TNF-α在胎膜早破早产孕妇血清、羊水中的表达及意义

目的研究IL-6、MMP-9、TNF-α在未足月胎膜早破早产孕妇的血清、羊水中的含量及表达,探讨其与胎膜早破早产的关系。方法采用酶联免疫吸附法检测30例胎膜早破早产孕妇（PPROM组）与20例正常孕妇（对照组）血清和羊水中的IL-6、MMP-9、TNF-α的含量,同时进行胎膜的病理检查。结果 PPROM组母血清及羊水中IL-6、MMP-9的含量均高于对照（P〈0.05）,羊水中TNF-α的含量较对照组高（P〈0.05）。PPROM组绒毛膜羊膜炎者血清、羊水中IL-6、TNF-α（P〈0.05）、MMP-9（P〈0.01）水平均高于非绒毛膜羊膜炎者。结论孕妇血清、羊水中IL-6、MMP-9、TNF-α水平与PPROM感染引起的早产有关,检测其水平可作为PPROM良好的预测指标。     

Regulation of matrix metalloproteinase-2(gelatinase A, MMP-2), membrane-type matrixmetalloproteinase-1 (MT1-MMP) and tissue inhibitorof metalloproteinases-2 (TIMP-2) expression byelastin-derived peptides in human HT-1080 fibrosarcoma cell line

Soluble kappa-elastin peptides were shown to stimulate the expression of MMP-2 (but not MMP-9) byhuman fibrosarcoma HT-1080 cells, both at the protein and mRNA levels; maximal effect being observedat a concentration of 25 mg/ml of kappa-elastin. The stimulatory effect could be reproduced using Val-Gly-Val-Ala-Pro-Gly (VGVAPG) peptide, an elastin-derived hydrophobic hexapeptide which represented theelastin receptor binding sequence of tropoelastin. Furthermore, treatment of cells with lactose (30 mM),which dissociated 67-kDa elastin binding protein (EBP) from cell surfaces, completely abolished this effect,suggesting that the elastin receptor could mediate such a response. Using a specific monoclonal antibody,67-kDa EBP was detected in HT-1080 membrane preparations by Western immunoblotting. Following treat-mentwith 25 mg/ml kappa-elastin or 200 mg/ml VGVAPG, increased levels of the active 62-kDa form ofMMP-2 were found in HT-1080 cell extracts. Stimulation of MT1-MMP mRNA expression by treatmentwith elastin-derived peptides (EDPs) was shown by competitive polymerase chain reaction (PCR). A reversezymography analysis revealed that EDPs also stimulated TIMP-2 (but not TIMP-1) production by HT-1080cells. Competitive PCR confirmed increased TIMP-2 mRNA expression by such treatment. These resultssuggest that occupancy of the 67-kDa elastin receptor by elastin-derived peptides enhanced both expressionand activation of proMMP-2 and consequently, could promote the invasive/metastatic ability of tumor cellsexpressing this receptor. ©Kluwer Academic Publishers     

High levels of MMP-2, MMP-9, MT1-MMP and TIMP-2 mRNA correlate with poor survival in ovarian carcinoma

The object of this study was to analyze the potential association between the expression of MMP-2, MMP-9, MT1-MMP and TIMP-2, and disease outcome in advanced-stage ovarian carcinomas. Sections from 70 paraffin-embedded blocks (36 primary ovarian carcinomas and 34 metastatic lesions) from 45 patients diagnosed with advanced stage ovarian carcinomas (FIGO stages III–IV) were studied using mRNA in situ hybridization (ISH) technique. Patients were divided retrospectively in two groups based on disease outcome. Long-term survivors (21 patients) and short-term survivors (24 patients) were defined using a double cut-off of 36 months for disease-free survival (DFS) and 60 months for overall survival (OS). Mean follow-up period for patients that were diagnosed with advanced-stage carcinoma was 70 months. The mean values for DFS and OS were 109 and 125 months for long-term survivors, as compared to 3 and 21 months for short-term survivors, respectively. Intense mRNA signals were detected more frequently in tumor cells of short-term survivors with use of all four probes. Comparable findings were observed in peritumoral stromal cells with ISH for MMP-2, MMP-9 and TIMP-2 mRNA. Notably, primary tumors with intense mRNA signal for TIMP-2 (No = 14) were uniformly associated with a fatal outcome. In univariate analysis of primary tumors, mRNA levels of TIMP-2 in stromal cells (P = 0.0002), as well as for MMP-9 (P = 0.012) and TIMP-2 (P = 0.02) in tumor cells, correlated with poor outcome. In univariate analysis of metastatic lesions, mRNA levels of TIMP-2 in stromal cells (P = 0.031), as well as for MMP-2 (P = 0.027) and MT1-MMP (P = 0.008) in tumor cells, correlated with poor outcome. Interestingly, the presence of MT1-MMP in stromal cells correlated with longer survival (P = 0.025). In a multivariate analysis of ISH results for primary tumors, TIMP-2 levels in stromal cells (P = 0.006) and MMP-9 levels in tumor cells (P = 0.011) retained their predictive value. We conclude that MMP-2, MMP-9, MT1-MMP and TIMP-2 are valid markers of poor survival in advanced-stage ovarian carcinoma. This revised version was published online in July 2006 with corrections to the Cover Date.     

Upregulated MT1-MMP/TIMP-2 axis in the TSU-Pr1-B1/B2 model of metastatic progression in transitional cell carcinoma of the bladder

Muscle invasive transitional cell carcinoma (TCC) of the bladder is associated with a high frequency of metastasis, resulting in poor prognosis for patients presenting with this disease. Models that capture and demonstrate step-wise enhancement of elements of the human metastatic cascade on a similar genetic background are useful research tools. We have utilized the transitional cell carcinoma cell line TSU-Pr1 to develop an in vivo experimental model of bladder TCC metastasis. TSU-Pr1 cells were inoculated into the left cardiac ventricle of SCID mice and the development of bone metastases was monitored using high resolution X-ray. Tumor tissue from a single bone lesion was excised and cultured in vitro to generate the TSU-Pr1-B1 subline. This cycle was repeated with the TSU-Pr1-B1 cells to generate the successive subline TSU-Pr1-B2. DNA profiling and karyotype analysis confirmed the genetic relationship of these three cell lines. In vitro, the growth rate of these cell lines was not significantly different. However, following intracardiac inoculation TSU-Pr1, TSU-Pr1-B1 and TSU-Pr1-B2 exhibited increasing metastatic potential with a concomitant decrease in time to the onset of radiologically detectable metastatic bone lesions. Significant elevations in the levels of mRNA expression of the matrix metalloproteases (MMPs) membrane type 1-MMP (MT1-MMP), MT2-MMP and MMP-9, and their inhibitor, tissue inhibitor of metalloprotease-2 (TIMP-2), across the progressively metastatic cell lines, were detected by quantitative PCR. Given the role of MT1-MMP and TIMP-2 in MMP-2 activation, and the upregulation of MMP-9, these data suggest an important role for matrix remodeling, particularly basement membrane, in this progression. The TSU-Pr1-B1/B2 model holds promise for further identification of important molecules.     

MT1-MMP correlates with MMP-2 activation potential seen after epithelial to mesenchymal transition in human breast carcinoma cells

We have previously reported that human breast carcinoma (HBC) cell lines expressing the mesenchymal intermediate filament protein vimentin (VIM+) are highly invasive in vitro, and highly metastatic in nude mice when compared to their VIM– counterparts. Since only VIM+ cell lines can be induced to activate matrix metalloproteinase-2 (MMP-2) upon stimulation with Concanavalin A (Con A), we have examined here membrane type 1 MMP (MT1-MMP), a cell surface activator of MMP-2. Northern analysis reveals baseline expression of MT1-MMP in five of the six VIM+ cell lines studied (MDA-MB-231, MDA-MB-435, BT-549, Hs578T, MCF-7ADR), each of which showed variable activation of exogenous MMP-2 after treatment with Con A. In contrast, the four VIM–, poorly invasive HBC cell lines studied (MCF-7, T47D, MDA-MB 468, ZR-75-1) lacked baseline MT1-MMP mRNA expression, and showed no induction of either MT1-MMP expression or MMP-2-activation with Con A. Such differential MT1-MMP expression was confirmed in vivo using in situ hybridization analysis of nude mouse tumor xenografts of representative cell lines. Western analysis of the MDA-MB-231 cells revealed baseline membrane expression of a 60 kDa species, which was strongly induced by Con A treatment along with a weaker band co-migrating with that from MT1-MMP-transfected COS-1 cells (63 kDa), presumably representing latent MT1-MMP. MT1-MMP immunofluorescence strongly decorated Con A-stimulated MDA-MB-231 cells in a manner consistent with membranous staining, but did not decorate the unstimulated MDA-MB-231 cells or MCF-7 cells under either condition. Collectively, the results suggest the constitutive production of active MT1-MMP which is unavailable for either MMP-2 activation or immuno-decoration until Con A treatment. Since VIM expression arises by virtue of the so-called epithelial to mesenchymal transition (EMT) in invasive embryonic epithelia, we propose that this represents a major metastasis mechanism in breast carcinomas. MT1-MMP on the surface of such ÔfibroblastoidÕ carcinoma cells may mediate a paracrine loop for the utilization of stromally produced MMP-2, and contribute to the poorer survival associated with VIM+ breast carcinomas.     

Impaired alveolization in mice deficient in membrane-type matrix metalloproteinase 1 (MT1-MMP)

To clarify the involvement of membrane-type matrix metalloproteinase 1 (MT1-MMP) in lung organogenesis, we studied the lung morphology of 13-day-old MT1-MMP null mice. The lung architecture in MT1-MMP null mice was abnormal, and the airspace compartments were characterized by smooth walls and larger size. Most of the compartment wall consisted of one or two layers of cells and interstitial connective tissue that was thicker than that of normal alveoli. The wall frequently had capillaries on both sides of the interstitial connective tissue. These findings indicate that the lung in MT1-MMP null mice at 13 days of age is comparable to that of neonatal mice, i.e., it represents the stage before alveolization, suggesting that the generation of a large respiratory surface – the final process of lung development – is impaired in MT1-MMP null mice. Moreover, a zymography assay revealed decreased activity of matrix metalloproteinase 2 (MMP-2) in MT1-MMP null mice, suggesting that activation of pro-MMP-2 by MT1-MMP is critical in this process.     

Soluble membrane-type 1 matrix metalloproteinase (MT1-MMP) and gelatinase A (MMP-2) in induced sputum and bronchoalveolar lavage fluid of human bronchial asthma and bronchiectasis

The aim of this study was to investigate the involvement of the MT1-MMP/MMP-2 cascade in induced sputum (IS) and bronchoalveolar lavage fluid (BALF) from bronchial asthma (BA) and bronchiectasis (BE) patients and healthy controls. The molecular forms and cellular origins of MT1-MMP and MMP-2 were determined by Western immunoblotting, immunohistochemistry and in situ hybridization. Elevated levels of soluble activated and autocatalyzed MT1-MMP species as well as activated forms of MMP-2 in IS and BALF samples from BA and BE patients were evidenced. The activation degrees of soluble MT1-MMP and MMP-2 were significantly correlated in BA and BE IS and BALF. Only low levels of both these MMPs were observed in healthy control IS and BALF. The co-expression of MMP-2 with MT1-MMP was evidenced by double immunostaining in bronchial epithelial cells, submucosal glandular cells, smooth muscle cells and monocyte/macrophages. The MT1-MMP/MMP-2 cascade is present and active in human inflammatory lung disease fluid and tissue samples. This cascade seemingly reflects the active destructive phases of these chronic lung diseases.     

MT2-MMP expression associates with tumor progression and angiogenesis in human lung cancer

Matrix metalloproteinases (MMPs) are a family of important proteolytic enzymes that play an important role in the remodeling of the tumor microenvironment and associate with tumorigenesis and metastasis. We previously reported that membrane type-2 MMP (MT2-MMP) is highly expressed in human esophageal cancer tissues, and its expression level is positively correlated to tumor size and intratumoral angiogenesis. In order to reveal whether MT2-MMP expression is operative in human lung cancer and its underlying physio-pathological role, in the present study, we examined both mRNA and protein expression levels of MT2-MMP in non-small cell lung caner (NSCLC) tissues and in adjacent normal tissues by using real-time RT-PCR and immunohistochemistry respectively, which showed that both MT2-MMP mRNA (P=0.0359) and protein (P<0.0001) expression levels were significantly increased in cancer tissues in contrast to adjacent normal tissues. Moreover, we also found that the MT2-MMP protein level in cancer tissues positively correlated to lymph node metastasis (P=0.0483), tumor stage (P=0.0483), intra-tumoral microvessel density (MVD) (P=0.0445). We have not found statistically significant correlation between MT2-MMP expression and patients’ prognoses, but we found that the patients with both higher MT2-MMP protein expression and higher intra-tumoral microvessel density showed better prognoses than that of the patients with either higher MT2-MMP protein expression or higher intra-tumoral microvessel density (P=0.0311). Thus, our data suggest that MT2-MMP expression positively involves in NSCLC, and might play an important role in promoting the tumor progression and intra-tumoral angiogenesis in NSCLC.     

非小细胞肺癌组织中MMP-9、TIMP-1表达及意义

目的 探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中基质金属蛋白酶9(MMP-9)、组织金属蛋白酶抑制剂-1(TIMP-1)的表达及其生物学行为的关系.方法 应用免疫组织化学SP法检测76例NSCLC和癌旁正常组织中MMP-9、TIMP-1的表达,并分析其表达与肺癌组织类型、肿瘤大小、TNM分期、分化程度、淋巴结转移的相关性.结果 MMP-9、TIMP-1在肺癌组织中的阳性表达率明显高于癌旁正常组织(P<0.05).NSCLC中MMP-9、TIMP-1表达与肿瘤的分化程度、临床分期、淋巴结转移有相关性(P<0.05),与肿瘤病理分型无关(P>0.05).结论 检测NSCLC中组织的MMP-9、TIMP-1的表达对判断肿瘤的恶性程度和预后评估有一定的意义.