siRNA介导S100A4基因沉默对胃癌细胞中MMP-2和E-cadherin表达及细胞侵袭能力的影响

目的观察用RNA干扰技术沉默人胃癌细胞株MGC-803中钙结合蛋白S100A4的基因,对基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)和E-钙黏素(E-cadherin)表达变化,探讨S100A4对胃癌细胞侵袭能力的影响及作用机制。方法采用RNA干扰技术将针对S100A4和非特异性阴性对照序列的si RNA转染至人胃癌MGC-803细胞,利用荧光实时定量RT-PCR和蛋白免疫印迹法(Western Blot)检测S100A4基因和蛋白的表达,筛选S100A4基因沉默组和阴性对照组细胞。采用Western Blot方法检测MMP-2和E-cadherin蛋白表达。利用Transwell Matrigel方法检测各实验组中MGC-803细胞体外侵袭能力。结果成功建立S100A4基因沉默的人胃癌MGC-803细胞。S100A4基因沉默的胃癌MGC-803细胞中MMP-2蛋白表达降低,而E-cadherin蛋白表达明显上调;细胞体外侵袭能力亦显著下降。结论 S100A4能通过调控MMP-2和E-cadherin的表达影响胃癌细胞MGC-803的侵袭能力。     

低氧抑制猪肺动脉平滑肌细胞MMP-2和MMP-9的表达

目的探讨低氧对猪肺动脉平滑肌细胞（PASMC）分泌基质金属蛋白酶（MMPs）的影响。方法采用酶谱法测定PASMC培养基中MMP-2和MMP-9的酶活性，免疫印迹法检测培养基中MMP-2和MMP-9的蛋白表达，免疫组化法测定细胞原位MMP-2和MMP-9的蛋白表达，RT-PCR法检测mRNA的表达。结果低氧后PASMC分泌的MMP-2酶活性、细胞内外蛋白表达量、mRNA表达量均下降；MMP-9酶活性、细胞外蛋白表达量下降，而细胞内蛋白表达无变化。结论低氧可抑制PASMC分泌MMP-2和MMP-9的酶活性，其机制可能是低氧影响PASMC中MMP-2基因的转录、影响MMP-9蛋白表达后的分泌与活化，导致MMP-2和MMP-9酶活性的改变。     

整合素αvβ6和MMP-9在胃癌中表达及其与胃癌病理生物学特征的关系

目的:探讨整合素αvβ6、MMP-9在胃癌中表达及其与胃癌病理生物学特征的关系。方法:采用EnVision免疫组化方法检测94例胃癌及癌旁胃黏膜组织中整合素αvβ6、MMP-9的表达情况,并分析两者表达与胃癌病理生物学特征的关系。结果:胃癌组织中整合素αvβ6和MMP-9阳性表达率明显高于癌旁胃黏膜组织(P<0.01)。整合素αvβ6在胃癌组织中的表达与Lauren分型、组织分化程度、是否有淋巴结转移(N分期)以及TNM分期密切相关(P<0.01),而MMP-9在胃癌组织中的表达与Lauren分型、组织分化程度、是否有淋巴结转移(N分期)、浸润深度以及TNM分期密切相关(P<0.01)。在胃癌组织中整合素αvβ6和MMP-9的表达呈正相关(r=0.672,P<0.01)。整合素αvβ6和MMP-9表达阴性患者的5年生存率明显优于整合素αvβ6和MMP-9表达阳性的患者(P<0.01)。Cox回归多因素分析表明Lauren分型、是否有淋巴结转移(N分期)、整合素αvβ6和MMP-9高表达是影响患者5年生存率的独立预后因素(P<0.05)。结论:整合素αvβ6和MMP-9的高表达与胃癌的浸润、转移密切相关;两者表达可作为判断胃癌患者预后的参考指标及治疗的靶向目标。     

慢性肾炎患者治疗前后血清MMP-2、MMP-9水平检测及其临床意义

文献报道,基质金属蛋白酶（MMPS）是一种具有共同生化活性可降解的细胞外基质（extra—cellular matrix,ECM）酶。MMP-2在未成熟的肾单位上皮中已有表达。MMP-9检测的临床意义远高于常规的尿微量白蛋白水平检测,在临床实践中有重要的临床价值。国内尚少见慢性肾炎患者治疗前后血清MMP-2和MMP-9联检的相关报导。为此,本文现将初步的探讨报告如下。     

肺腺癌组织中肿瘤相关中性粒细胞浸润和MMP-9的表达及其临床病理意义

目的 观察肺腺癌中肿瘤相关中性粒细胞(tumor associated neutrophils,TANs)的浸润及其与MMP-9表达的相关性,探讨其临床病理学意义.方法 收集肺腺癌手术切除组织100例,采用免疫组化法检测肺腺癌中TANs浸润密度及MMP-9表达,分析其与临床病理特征的关系及意义.利用生物信息学技术分析肺...     

慢性肾炎患者输注红细胞前后血清Ferritin、MMP-2和MMP-9检测的临床意义

目的：探讨了慢性肾炎患者在输注红细胞前后Ferritin、MMP-2和MMP-9水平的变化。方法：应用放射免疫分析和酶联法对32例慢性肾炎患者进行了输注红细胞前后Ferritin、MMP-2和MMP-9检测,并与35名正常人组作比较。结果：慢性肾炎患者在输注红细胞前血清Ferritin和MMP-9水平非常显著地低于正常人组（P〈0.01）,而MMP-2水平则显著地高于正常人组（P〈0.01）。经2周输注红细胞后则与正常人组比较无显著性差异（P〉0.05）。结论：检测慢性肾炎患者输注红细胞治疗前后血清Ferritin、MMP-2和MMP-9水平的变化对其病情的发展和预后的判断均具有重要的临床价值。     

前列腺间质细胞中雌二醇对MMP-2，MMP-9及其组织抑制因子表达的影响

目的： 研究雌二醇(E2)对前列腺间质细胞中基质金属蛋白酶(MMP-2、MMP-9）及其组织抑制因子1(TIMP-1)、TIMP-2和雌激素受体α、β（ERα、ERβ）的影响。方法： 实时定量PCR法检测E2在前列腺间质细胞中对MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA水平的影响；半定量RT-PCR法检测E2对ERα、ERβ mRNA水平的影响。酶谱电泳法检测MMP-2、MMP-9的活性。Western blotting检测E2在前列腺间质细胞中对ERα蛋白水平的影响。结果： 前列腺间质细胞中有MMP-2和ERα mRNA的表达，未检测到MMP-9 mRNA；培养液中检测到MMP-2前体（pro-MMP-2），未检测到其活性形式，也未检测到MMP-9前体及其活性形式。用E2处理间质细胞后MMP-2 mRNA水平降低，pro-MMP-2蛋白量减少，雌激素受体抑制剂ICI 182.780可抑制此作用。E2对TIMP-1,2 mRNA表达无显著影响。E2能够增加前列腺间质细胞中ERα mRNA及其蛋白表达的水平。结论： E2能够通过ERα下调前列腺间质细胞中MMP-2的表达。     

慢性肾炎患者SF、血清CysC和MMP-2、MMP-9检测的临床意义

目的:探索了慢性肾炎患者治疗前后SF、血清CysC和MMP-2、MMP-9水平的变化及临床意义。方法:应用放射免疫分析和酶联法对31例慢性肾炎患者进行了治疗前后SF、血清CysC和MMP-2、MMP-9检测,并与35名正常人作比较。结果:慢性肾炎患者在治疗前SF、血清MMP-9水平非常显著地低于正常人组(P<0.01),而CysC、MMP-2水平又非常显著地高于正常人组(P<0.01),经中西医结合治疗3个月后与正常人组比较仍有显著性差异(P<0.05)。结论:检测慢性肾炎患者SF、血清CysC和MMP-2、MMP-9水平的变化对其病情的发展和预后的判断均有重要的临床价值。     

咪达普利拉对人心脏成纤维细胞基质金属蛋白酶-2和抑制剂-2的作用及机制研究

目的：研究血管紧张素转化酶抑制剂（angiotensin converting enzyme inhibitor，ACEI）咪达普利活性代谢物咪达普利拉对白细胞介素－1β（interleukin -1β，IL-1β）诱导的心脏成纤维细胞基质金属蛋白酶（matrix metalloproteinases，MMPs）和基质金属蛋白酶2型抑制剂（type 2 tissue inhibitor of matrix metalloproteinase，TIMP-2）表达的影响及其可能机制。方法:原代人心脏成纤维细胞从美国细胞应用所购买。用RT-PCR检测心脏成纤维细胞MMP-2、MMP-9和TIMP-2 mRNA水平的变化；用凝胶酶谱法分析心脏成纤维细胞中MMP-2、MMP-9的活性；用Griess方法检测培养上清一氧化氮（nitric oxide，NO）水平。结果:通过凝胶酶谱分析，RT-PCR和Griess方法发现IL-1β能显著增加MMP-2基因转录以及活性(P<0.05)，同时也显著增加了细胞培养上清中NO的产生(P<0.05)。IL-1β的这些效应可以被咪达普利拉和外源性NO合酶抑制剂L-单甲基精氨酸（NG-methyl L-Arginine，L-NMMA）所抑制(P<0.05)，而外源性NO供体亚硝基铁氰化钠(sodium nitroprusside，SNP）可以取消咪达普利拉对于NO、MMP-2的有效作用(P<0.05)。实验中发现IL-1β以及咪达普利拉对TIMP-2基因转录无明显作用(P>0.05)。结论:咪达普利拉能通过NO途径抑制IL-1β诱导的心脏成纤维细胞MMP-2的合成和活性；提示ACEI抗心肌重塑以及心力衰竭的部分作用可能是通过MMPs途径实现的。     

可溶性趋化因子对人单核细胞表达及基质金属蛋白酶-9的影响

目的观察趋化因子Fractalkine（CX3CL1,Fkn）对人单核细胞株U937细胞表达与分泌基质金属蛋白酶-9（matrix metalloproteinase-9,MMP-9）的影响。方法使用不同浓度的人重组Fkn干预U937细胞,RT-PCR法检测细胞MMP-9基因表达,明胶酶谱法检测细胞上清中MMP-9水平。结果不同浓度人重组Fkn干预U937细胞MMP-9基因表达及上清中MMP-9水平低于空白对照组。结论Fkn可抑制U937细胞表达与分泌MMP-9。     

基质金属蛋白酶在肿瘤发展过程中保护作用的研究进展

基质金属蛋白酶(MMPs)家族是细胞外蛋白水解酶,几乎可以降解所有细胞外的基质成份.普遍认为肿瘤细胞或周围基质中MMPs特异性高表达是肿瘤形成的一个重要原因,因此研究用人工合成的MMP抑制剂治疗肿瘤已成为研究领域的热点.遗憾的是在临床试验阶段该治疗方法并没有达到预期的效果.近年来通过对肿瘤初期、转移后的表达研究发现,MMPs在肿瘤发展的多个阶段起着抑制肿瘤生长的作用.     

A prognostic index in skin melanoma through the combination of matrix metalloproteinase-2, Ki67, and p53

The objective of this immunohistochemical study was to explore the roles of Ki67 and p53 in conjunction with matrix metalloproteinase-2 and matrix metalloproteinase-9, tissue inhibitors of metalloproteinase-1 and tissue inhibitors of metalloproteinase-2 in a series of 157 cases of skin melanomas. Elevated Ki67 expression and positive staining for p53 correlated to the propensity to metastasize (P = .016) and to declined disease-specific survival, as well as to shortened recurrence-free survival. In patients with a high immunoreaction for Ki67, the 10-year disease-specific survival was 39% compared with 73% in patients with a low Ki67 expression (P = .03). In cases with a positive p53 expression in melanoma cells, the 10-year disease-specific survival was 59% compared with 76% in patients with a negative immunoreaction for p53 (P = .005). Overexpression of the matrix metalloproteinase 2 protein in conjunction with overexpression of Ki67 characterized melanomas with high metastatic potential and was associated with declined survival with a 10-year disease-specific survival of 33% compared with 85% in the cases with low matrix metalloproteinase-2 and low Ki-67 levels (P = .002). Similarly, in cases with overexpression of matrix metalloproteinase-2 and a positive immunoreaction for p53, the 10-year disease-specific survival was only 42% compared with 80% in patients with matrix metalloproteinase-2 less than 20% and a negative immunostaining for p53 (P < .001). The presence of all 3 adverse prognostic factors was prognostically more significant than any marker alone with a 10-year survival of only 28%. This combination of determining matrix metalloproteinase 2, Ki67, and p53 immunoreactive proteins could be beneficial in the selection of high-risk melanoma patients for future adjuvant trials.     

基质金属蛋白酶-2、3及其组织抑制剂TIMP-1在子宫内膜异位症中的表达

目的探讨基质金属蛋白酶(MMP-2、MMP-3)及其抑制剂(TIMP-1)在子宫内膜异位症发生及发展中的作用.方法采用免疫组化SP法分别测定MMP-2、MMP-3 、TIMP-1在卵巢子宫内膜异位症异位内膜60例(A组),子宫腺肌症异位内膜40例(B组),子宫肌瘤子宫内膜30例(对照组C)的表达强度.结果 A、B组中MMP-2、MMP-3的表达强度明显高于对照组(P<0.05)而TIMP-1的表达明显低于对照组(P<0.05);A、B组间MMP-2、MMP-3 、TIMP-1 的表达无明显差异(P>0.05).结论在子宫内膜异位症中MMP-2、MMP-3的过度表达及TIMP-1的低表达可能与内异症的发生与发展有关.     

Doxycycline对PC—3细胞金属蛋白酶表达及生物学行为影响的意义

目的 研究Doxycycline抑制雄性激素非依赖型前列腺癌细胞PC－3细胞金属蛋白酶蛋白酶的表达及其与体外侵袭转移能力的关系。方法 以免疫组织化学方法和transwell小室法研究不同浓度的Doxycycline对PC-3金属蛋白酶的表达的影响及对体外侵袭转移能力的影响。结果 免疫组织化学方法检测结果表明Doxycycline可以抑制PC－3细胞对金属蛋白酶蛋白的表达,transwell小室法结果显示Doxycycline可以抑制PC－3的侵袭转移能力,且两者均具有浓度依赖性。结论 Doxycycline可以抑制PC－3的侵袭及转移,与其抑制金属蛋白酶的表达有关。为Doxycycline治疗前列腺癌提供了实验依据。     

基质金属蛋白酶的结构、功能和调节

基质金属蛋白酶（MMPs)是一个蛋白水解酶大家族，可降解细胞外很多种基质成分。根据体外底物专一性，MMPs可分为胶原酶，明胶酶，基质降解素以及膜型MMPs等，越来越多的研究表明，MMPs在肿瘤侵袭转移中起着重要的作用，而且此作用不仅仅限于它有利于细胞外基质的降解，还对肿瘤微环境的维持和促进肿瘤生长起着重要作用，本文对MMPs的结构，功能及其调节进行综述。     

Reversion-inducing cysteine-rich protein with Kazal motif (RECK) expression: an independent prognostic marker of survival in colorectal cancer

Patient prognosis in colorectal cancer is determined as in most solid cancers by the extent of local invasion and the presence of lymph node and distant metastases. The invasive potential of a tumor depends on the ability to degrade extracellular matrix proteins, for example, by matrix metalloproteinases. An important inhibitor of matrix metalloproteinases is reversion-inducing cysteine-rich protein with Kazal motifs (RECK), a membrane-anchored glycoprotein. This study investigated the prognostic relevance of RECK expression in colorectal cancer in a cohort of 283 patients. Analysis of immunohistochemical tissue microarray data showed that RECK protein levels did not seem to correlate with clinicopathologic parameters (Spearman rank correlation coefficients between -0.14 and -0.18) and that decreased RECK expression was an independent prognostic factor of poor survival, with a mean survival of 70 months in RECK-negative (146 cases) versus 97 months in RECK-positive patients (137 cases) (log-rank test, P = .002).     

Expression of matrix metalloproteinases and their tissue inhibitors in the serum and cerebrospinal fluid of patients with meningitis

Meningitis is associated with an imbalance between matrix metalloproteinases (MMPs) and endogenous tissue inhibitors of MMP (TIMPs). Serum and CSF were collected prospectively from all patients with meningitis between January 2008 and December 2008 to measure the concentrations of MMP/TIMP in those patients who underwent a lumbar puncture for a presumptive diagnosis of meningitis. A total of 199 patients were enrolled into the study. The concentrations of CSF MMP‐9 and TIMP‐1 were significantly higher in the meningitis group compared with the control group (p 0.032 and p <0.001, respectively). However, the CSF TIMP‐4 levels were significantly lower in the meningitis groups compared with the control groups (p <0.001). Patients with bacterial meningitis had higher CSF MMP‐9 and TIMP‐1 levels than those who had aseptic meningitis and controls. Patients with various infectious meningitis etiologies tended to have higher CSF MMP‐9 expression by gelatin zymography when compared with the controls. In conclusion, MMP/TIMP system dysregulation was found in patients with meningitis, and CSF MMP and TIMP might act as novel indicators in patients with meningitis.     

基质金属蛋白酶在多发性硬化发病中的作用机制及治疗现状

多发性硬化（MS）是中枢神经系统炎性脱髓鞘疾病,基质金属蛋白酶是一组蛋白水解酶,有重要的病理和生理作用,近年的研究表明它在（MS）的发生和疾病进展中都具有重要作用,参与主要的发病过程,具有多种发病机制,抑制其活性具有有效的治疗意义。     

Elevation of ADAM10, ADAM17, MMP-2 and MMP-9 expression with media degeneration features CaCl2-induced thoracic aortic aneurysm in a rat model

Purpose

This study was designed to establish a rat model of thoracic aortic aneurysm (TAA) by calcium chloride (CaCl₂)-induced arterial injury and to explore the potential role of a disintegrin and metalloproteinase (ADAM), matrix metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs) in TAA formation.

Methods

Thoracic aorta of male Sprague-Dawley rats was exposed to 0.5 M CaCl₂ or normal saline (NaCl). After 12 weeks, animals were euthanized, and CaCl₂-treated, CaCl₂-untreated (n = 12) and NaCl-treated aortic segments (n = 12) were collected for histological and molecular assessments. MMP-TIMP and ADAM mRNAs were semi-quantitatively analyzed and protein expressions were determined by immunohistochemistry.

Results

Despite similar external diameters among CaCl₂-treated, non-CaCl₂-treated and NaCl-treated segments, aneurymal alteration (n = 6, 50%), media degeneration with regional disruption, fragmentation of elastic fiber, and increased collagen deposition (n = 12, 100%) were demonstrated in CaCl₂-treated segments. MMP-2, MMP-9, ADAM-10 and ADAM-17 mRNA levels were increased in CaCl₂-treated segments (all p < 0.01), with trends of elevation in CaCl₂-untreated segments, as compared with NaCl-treated segments. Immunohistochemistry displayed significantly increased expressions of MMP-2, MMP-9, ADAM-10 and ADAM-17 (all p < 0.01) in intima and media for CaCl₂-treated segments. TIMP mRNA and tissue levels did not differ obviously among the three aortic segments.

Conclusion

This study establishes a TAA model by periarterial CaCl₂ exposure in rats, and demonstrates a significant elevation of expression of MMP-2, MMP-9, ADAM10 and ADAM17 in the pathogenesis of vascular remodeling.     

Genetic variations in matrix metalloproteinases may be associated with increased risk of ulcerative colitis

Increased production of matrix metalloproteinases (MMPs) plays an important role in tissue damage in inflammatory bowel disease (IBD). Genetically encoded variation between individuals in MMP production may therefore contribute to disease onset, type, or severity. We undertook an extensive candidate gene single nucleotide polymorphism (SNP) study of MMP-1, -2, -3, -7, -8, -9, -10, -12, -13, and -14 and tissue inhibitor of metalloproteinases (TIMPs)-1, -3, and -4 in ulcerative colitis (UC). We identified tagging SNPs across these genes, and genotyped these SNPs in a Caucasian New Zealand dataset consisting of 419 UC patients and 907 controls. SNPs in a number of MMP genes were associated with UC. After correcting for multiple testing SNPs in MMP-3, MMP-8, MMP-10, and MMP-14 remained significant in their associations with UC. In a second study, using samples from a Dutch cohort, most of the significant findings in the New Zealand cohort were not replicated. However, data from an international meta-analysis provide some support for the initial findings. In conclusion, this study provides preliminary evidence to suggest that genetic variation in the MMPs may play a role in interindividual differences in UC susceptibility and clinical outcome. Further studies are needed in other cohorts to determine the robustness of these observations in different populations.