基质金属蛋白酶-2及其组织抑制因子在大肠癌组织中表达及临床意义

目的 检测基质金属蛋白酶-2(MMP-2)及其组织抑制因子(TIMP-2)在大肠癌组织中的表达,探讨其临床意义.方法 采用免疫组织化学方法 、Western blot及实时定量PCR法检测50例大肠癌组织及其邻近的癌旁组织、50例正常大肠组织中MMP-2和TIMP-2的表达,并分析大肠癌组织中MMP-2和TIMP-2的表达与病理分级、临床分期及转移的关系.结果 大肠癌MMP-2蛋白阳性表达率为72.0%,显著高于癌旁组织的40.0%,正常组织的20.0%,呈递减趋势(P<0.01).TIMP-2蛋白阳性表达率在大肠癌为36.0%,显著低于癌旁组织的62.0%和正常组织的80.0%,呈递增趋势(P<0.01).Western blot结果 与免疫组织化学一致,灰度分析结果 显示差异有统计学意义(P<0.01).实时定量PCR结果 显示大肠癌MMP-2和TIMP-2扩增倍数值为(2.49±0.48)、(0.34±0.18),癌旁组织为(1.47±0.23)、(0.63±0.21),两组表达差异有统计学意义(P<0.01).大肠癌MMP-2的高表达和肿瘤分化程度、TNM分期、淋巴结转移呈正相关,而TIMP-2却相反.结论 大肠癌组织中MMP-2的高表达和TIMP-2的低表达,提示MMP-2和TIMP-2两者之间的平衡失调可能是肿瘤侵袭、转移的最终机制.     

基质金属蛋白酶-2和CD147在膀胱癌的表达及临床意义

目的 通过检测基质金属蛋白酶-2(MMP-2)和CD147在膀胱移行细胞癌中的表达,探讨其在浸润转移中的作用。方法 用免疫组织化学链霉素抗生物素蛋白-过氧化酶法检测45例膀胱癌组织和9例正常膀胱粘膜中MMP-2及CD147的表达,并与临床病理指标进行对照分析。结果 膀胱癌组织中 MMP-2和 CD147过度表达,阳性率分别为 78％和 73％,MMP-2以浸润前缘的间质细胞表达为主,CD147表达于癌细胞。两者的联合表达率为66.7％,r=0.605(P<0.01)。结论 MMP-2和CD147与膀胱癌的恶性表型密切相关,间质细胞来源的MMP-2可能在膀胱癌的浸润转移中起主要作用。CD147与MMP-2的产生呈正相关,是膀胱癌潜在的治疗靶点。     

基质金属蛋白酶-1、金属蛋白酶组织抑制剂-1、增殖细胞核抗原在病理性瘢痕中的表达

目的：研究基质金属蛋白酶-1（MMP-1）、金属蛋白酶组织抑制剂-1（TIMP-1）、增殖细胞核抗原（PCNA）在病理性瘢痕中的表达及意义.方法：对58例病理性瘢痕手术切除标本采用免疫组化方法.结果：MMP-1、TIMP-1、PCNA在病理性瘢痕中呈阳性表达.结论：病理性瘢痕中细外基质合成增加为成纤维细胞过度增殖所致;病理性瘢痕形成与MMP-1、TIMP-1相互作用失衡有关.     

子宫内膜癌组织中基质金属蛋白酶-7及其组织抑制剂-2的表达

目的探讨子宫内膜癌(EC)组织中基质金属蛋白酶-7(MMP-7)及其组织抑制剂-2(TIMP--2)的表达,分析其与EC侵袭、转移的关系。方法选取64例EC组织标本及同期子宫内膜不典型增生组织标本20例、正常子宫内膜20例为对照组。用免疫组织化学方法检测MMP-7、TIMP-2的表达。结果 MMP-7在EC中阳性表达强度显著高于正常子宫内膜(P0.05);与不典型增生组比较差异无统计学意义(P0.05);MMP-7的阳性表达强度在不典型增生组高于正常内膜组(P0.05);MMP-7与手术-病理分期、肌层浸润深度、组织学分级及淋巴结的转移相关(P0.05)。TIMP-2在EC中阳性表达强度显著高于正常子宫内膜组和不典型增生组(P0.05);正常子宫内膜组和不典型增生组比较TIMP-2的表达无统计学意义(P0.05);TIMP-2仅与有无淋巴结转移有关(P0.05)。MMP-7和TIMP-2在EC组织中的表达呈正相关性(r=0.654,P0.001)。结论 MMP-7与TIMP-2在EC组织中均呈高表达;且子宫内膜不典型增生组MMP-7的表达强度明显高于正常内膜组,可能与EC前病变以及EC的发生发展、浸润和转移相关。     

基质金属蛋白酶-9及金属蛋白酶组织抑制剂-1对甲状腺乳头状癌诊断的意义

目的:探讨基质金属蛋白酶-9(MMP-9)和金属蛋白酶组织抑制剂-1(TIMP-1)在甲状腺肿瘤组织中的表达。方法:制备组织芯片,采用免疫组织化学和原位杂交技术检测56例甲状腺癌组织、56例癌旁组织和40例良性甲状腺病变组织中MMP-9和TIMP-1的表达情况。结果:MMP-9和TIMP-1蛋白在甲状腺癌组织的阳性表达率为71.4%和57.1%,MMP-9 mRNA和TIMP-1 mRNA在甲状腺癌组织中阳性表达率为67.9%和62.5%,均明显高于癌旁和良性甲状腺病变组织(P<0.05)。在甲状腺癌组织中,MMP-9、TIMP-1蛋白和MMP-9、TIMP-1 mRNA的表达,分别呈负相关性(RS=-0.309、-0.264,P<0.05)。结论:MMP-9和TIMP-1在组织中的检测有助于甲状腺癌的判断,并可作为预后评估的重要参考。     

胆囊癌中CD44v6和基质金属蛋白酶-2的表达及其临床意义

目的: 探讨胆囊癌中粘附分子CD44v6、基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)的表达及临床意义,以及二者之间的关系. 方法: 采用免疫组织化学方法检测42例胆囊癌、10例慢性结石性胆囊炎组织中CD44v6和MMP-2的表达. 结果: ①CD44v6阳性表达率在胆囊癌组为64.3%(27/42),明显高于慢性结石性胆囊炎组(0/10)(P<0.005);CD44v6表达与胆囊癌的组织类型、病理分级以及转移相关(P<0.05);CD44v6阳性的胆囊癌病人预后较差.②MMP-2在胆囊癌组织中表达率为61.9%,明显高于慢性结石性胆囊炎组(10%,P<0.005);MMP-2的表达与胆囊癌细胞的分化、转移和预后有关(P<0.05),与组织类型无关.③CD44v6和MMP-2在胆囊癌组织中表达呈正相关(P<0.005). 结论: CD44v6和MMP-2的表达与胆囊癌分化、转移、预后有关,在胆囊癌细胞的侵袭和转移中起重要作用.联合检测有利于评估胆囊癌的生物学行为和判断预后.     

胃癌基质金属蛋白酶-9表达的临床意义

目的探讨基质金属蛋白酶-9(MMP-9)表达与胃癌侵袭转移及预后的关系.方法采用SP免疫组织化学方法,检测20例正常胃黏膜上皮、20例异型增生、108例胃癌组织及47例相应癌转移淋巴结MMP-9的表达情况.结果正常胃黏膜和异型增生MMP-9阳性表达率分别为20.0%(4/20)和70%(14/20),均呈弱阳性表达胃癌组织MMP-9表达阳性率为88.9%(96/108),显著高于前两者(P＜0.05);癌转移淋巴结MMP-9阳性表达率为100%(47/47),明显高于胃癌原发灶(P＜0.05);MMP-9表达强度与胃癌细胞分化程度无明显相关(P＞0.05),与胃癌生长方式、浸润深度、淋巴结转移、远处转移、脉管侵犯及病期密切相关(均P＜0.05);MMP-9强阳性表达胃癌患者术后5年生存率明显下降(P＜0.05).结论MMP-9表达与胃癌侵袭转移及预后密切相关.     

基质金属蛋白酶抑制剂-2在贲门癌患者血清和肿瘤组织中的表达及其临床意义

我们应用酶联吸附免疫和免疫组织化学染色分别检测了40例贲门癌患者的术前血清和术后标本中基质金属蛋白酶抑制剂(TIMP)-2的表达,探讨TIMP-2在贲门癌发生发展中的作用,以及TIMP-2在术前血清和术后标本中表达的关系.     

尿液中组织基质金属蛋白酶抑制剂对筛查膀胱癌的意义

目的 初步探讨一种新的无创伤的筛查膀胱癌的方法。方法 收集50例膀胱癌患者术前（17例Ta-T1,21例T2,10例T3,2例T4;11例I级,23例Ⅱ例,16例Ⅲ级和11例正常人尿液,采用Western blot半定量方法检测其中基质金属蛋白酶-2（MMP-2）、MMP-9和组织基质金属蛋白酶抑制剂-2（TMP）的表达。结果 （1）正常人尿液中无MMP-2、MMP-9的表达,而TIMP-2表达阳性率为9.09%;（2）膀胱癌患者尿液中MMP-2、MMP-9、TIMP-2的阳性表达率分别为18%,12%,80%,且MMP-9的阳性率表达与肿瘤的病理分期和分级具有相关性;（3）半定量分析,MMP-2、MMP-9表达强度与患者分期分级无关,而在Ⅲ级和T4期患者尿液中TIMP-2表达平均强度具有高于I-Ⅱ级和Ta-T3期患者的趋势。结论 MMP-9在膀胱癌的侵袭过程中起着重要作用,通过检测尿液中TIMP-2的表达,可为临床诊断膀胱癌提供一种新的无创的初筛方法。     

CD147与基质金属蛋白酶-2、血管内皮生长因子在人膀胱移行细胞癌中的表达及其临床意义

目的 探讨膀胱移行细胞癌（BTCC）组织中细胞外基质金属蛋白酶诱导因子（CD147）与基质金属蛋白酶-2（MMP-2）、血管内皮生长因子（VEGF）的表达及其临床意义。方法采用免疫组织化学链霉菌抗生物素．蛋白过氧化酶（SP）法检测72例BTCC组织和12例正常膀胱组织CD147与MMP-2、VEGF的表达，分析CD147和MMP-2、VEGF间及其与BTCC部分临床生物学特性的相关性。结果 BTCC组织中有CD147、MMP-2、VEGF表达，其阳性率分别为68．1％、76．4％、70．8％。CD147表达与病理分级显著相关，与临床分期未见相关；MMP及VEGF表达与病理分级和临床分期显著相关；CD147表达与MMP-2、VEGF表达呈显著正相关（r=0．558，P〈0．01；r=0．406，P〈0．01）。结论 CD147在BTCC的浸润与转移中起重要作用，是新的治疗靶点。     

血清基质金属蛋白酶2和抑制因子与绝经后骨质疏松症的相关性

目的探讨绝经后妇女血清基质金属蛋白酶2（MMP-2）和抑制因子（TIMP-2）水平及其与绝经骨质疏松症指标的关系。方法将202名48～65岁绝经后妇女分为正常组、低骨量组和骨质疏松组，用酶联免疫吸附试验（EIJSA）测定的血清MMP-2、TIMP-2以及骨保护蛋白（OPG）、骨保护蛋白配体（OPGL），计算MMP-2／TIMP-2和OPG／OPGL比值，用双能X线吸收法（DEXA）测定腰椎正位、股骨颈、华氏区和大粗隆的骨密度（BMD）。结果①骨质疏松组中血清MMP-2的数值（1392±121）μg／L高于正常组（1123±141）μg／L（P〈0．05），而TIMP-2的数值（44．3±36．2）ng/ml低于正常组（47．8±30．2）ng/ml。②骨质疏松组中血清MMP-2和MMP-2／TIMP-2比值与骨密度、血精OPGL数值存在明显负相关性（P〈0．05），和OPG和OPG／OPGL比值存在明显正相关性（P〈0．05），TIMP-2和华氏区骨密度和OPG存在明显正相关性（P〈0．05）。结论血清MMP-2和MMP-2／TIMP-2比值与绝经后骨质疏松症妇女骨密度和骨代谢指标OPG、OPGL和OPG／OPGL比值具有关联性。血清MMP-2水平升高和MMP-2／TIMP-2比值降低可能为绝经后骨质疏松症伴随骨代谢转换过程增快的表现。     

乳腺癌中HER2基质金属蛋白酶-2和9的表达及其相互关系

目的　研究乳腺癌中HER2基因、基质金属蛋白酶 (MMP) 2、基质金属蛋白酶(MMP) 9的表达情况、与临床病理指标之间的关系以及它们之间的相互关系。方法　采用免疫组织化学的方法对 114例乳腺癌组织标本中HER2、MMP 2、MMP 9的表达情况进行检测。结果　乳腺癌组织中HER2、MMP 2、MMP 9的表达阳性率分别是 46.49%、78.95 %、68.42 %。HER2表达与淋巴结转移相关。原发肿瘤 >2cm或有淋巴结转移的患者中 ,其MMP 2、MMP 9表达明显高于原发肿瘤≤ 2cm或无淋巴结转移的患者 (P <0 .0 5 ) ,且MMP 2表达与临床分期相关 (P <0 .0 5 )。HER2表达与MMP 2、MMP 9表达相关 (P <0 .0 5 )。结论　HER2、MMP 2、MMP 9的阳性表达提示乳腺癌有较强的浸润转移能力 ,这 3种蛋白的表达在乳腺癌浸润转移过程中可能起协同作用。     

Localization and quantification of mRNA for matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in human benign and malignant prostatic tissue

BACKGROUND: The family of matrix metalloproteinases (MMPs) has been shown to be involved in proteolytic degradation of the extracellular matrix, which is an essential step in tumor invasion and metastasis. MMPs are tightly regulated by the levels of active enzymes and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). MMP-2 and its ratio to TIMP-2 have been associated with tumor recurrence and progression in a number of human malignancies. METHODS: We examined the relationship between MMP-2 and TIMP-2 mRNA expression in 42 men with malignant (n = 32) and benign (n = 10) prostates using nonisotopic in situ hybridization and Northern blot analysis. RESULTS: mRNA for MMP-2 and TIMP-2 was localized to the malignant epithelial cells of both high- and low-grade tumors in the periphery of the glands and in areas of extracapsular involvement, and to the glandular epithelium in the benign prostates. Using Northern blot analysis, the mean MMP-2 to TIMP-2 ratio was approximately one in the benign prostates and low-grade and -stage cancers. The MMP-2 to TIMP-2 ratio increased to 3.3 in the high-grade and 2.8 in the high-stage tumors. CONCLUSIONS: The results suggest a close association between MMP-2/TIMP-2 expression and local tumor invasion, with a disruption in expression of the two genes leading to disease progression. Future studies should focus on the activity of these enzymes and on the ratio of enzyme/inhibitor expression, which may become a useful prognostic marker in prostate cancer.     

罗格列酮对肝星状细胞基质金属蛋白酶-2和-9基因表达的影响

目的 观察过氧化物酶体增殖物活化受体γ(PPARγ)特异配体罗格列酮对肝星状细胞(HSC)表达基质金属蛋白酶(MMP)-2和MMP-9的影响.方法 在培养的HSC株中加入不同浓度的罗格列酮(终质量浓度为5、10、15、20 μmol/L),于24 h后收取细胞,利用半定量逆转录-聚合酶链反应(RT-PCR)测定MMP-2、MMP-9 mRNA表达.结果 MMP-2表达水平在罗格列酮5、10、15、20 μmol/L组分别为0.5708±0.0609、0.8900±0.0823、1.1348±0.1205、1.4490±0.0832,而空白对照组为0.3237±0.0796.MMP-9表达水平在罗格列酮5、10、15、20 μmol/L组分别为0.5487±0.0770、0.7554±0.0510、0.9497±0.0451、1.1088±0.0777,空白对照组为0.3220±0.0592.结论 罗格列酮可增强HSC对MMP-2、MMP-9的表达,并在一定范围内,呈剂量依赖性.     

An imbalance between matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 contributes to the development of early diabetic nephropathy.

BACKGROUND: High glucose and angiotensin-II (Ang-II) levels are the known important mediators of diabetic nephropathy. However, the effects of these mediators on matrix metalloproteinase-2 (MMP-2) and on tissue inhibitor of metalloproteinase-2 (TIMP-2) in proximal tubule cells have yet to be fully examined within the context of early stage diabetic nephropathy. METHODS: In this study, we attempted to characterize changes in MMP-2 and TIMP-2 in streptozotocin-induced diabetic rats. To further examine the molecular mechanisms involved, we evaluated the effects of high glucose (30 mM) or Ang-II on MMP-2, TIMP-2 and collagen synthesis in proximal tubule cells, and investigated whether MMP-2 and TIMP-2 are regulated via the TGF-beta1 pathway. RESULTS: In streptozotocin-induced diabetic rats, TIMP-2 mRNA and protein levels were significantly higher than in controls. Urinary protein excretion also showed a significant positive correlation with glomerular and tubular TIMP-2 protein expressions, and a negative correlation with MMP-2 expression. In cultured cells, both high glucose and Ang-II induced significant increases in TGF-beta1, TIMP-2, and in collagen synthesis, and significant decreases in MMP-2 gene expression and activity, and thus disrupted the balance between MMP-2 and TIMP-2. Moreover, treatment with a selective angiotensin type 1 (AT1) receptor antagonist significantly inhibited Ang-II mediated changes in TGF-beta1, MMP-2, TIMP-2, and in collagen production, suggesting the role of the AT1 receptor. The addition of exogenous TGF-beta1 produced an effect similar to those of high glucose and Ang-II. Furthermore, the inhibition of TGF-beta1 protein prevented Ang-II-induced MMP-2 and TIMP-2 alterations, suggesting the involvement of a TGF-beta1 pathway. CONCLUSIONS: High glucose or Ang-II treatment induce alterations in MMP-2 and TIMP-2 balance, which favour TIMP-2 over-activity. Moreover, Ang-II-mediated changes in the productions of MMP-2 and TIMP-2 occur via AT1 receptors and a TGF-beta1-dependent mechanism. These results suggest that an imbalance between the MMP-2 and TIMP-2, caused primarily by an increase in TIMP-2 activity, contributes to the pathogenesis of diabetic nephropathy.     

Connective tissue growth factor increases matrix metalloproteinase-2 and suppresses tissue inhibitor of matrix metalloproteinase-2 production by cultured renal interstitial fibroblasts

The involvement of gelatinase (matrix metalloproteinase-2 [MMP-2] and MMP-9) in the matrix remodeling and development of tubulointerstitial fibrosis has been studied recently, but relatively little is known about the regulators and the mechanisms controlling the activation and expression of gelatinase in renal fibroblasts. In these studies, the production and underlying signaling pathway for gelatinase by exogenous connective tissue growth factor (CTGF) treatment were investigated. Here, we show that CTGF acts as a potent promoter of the activation and expression of MMP-2, but not MMP-9 in normal rat kidney fibroblasts cell line (NRK-49F). We found that CTGF significantly increased the activity of MMP-2, as well as MMP-2 protein in conditioned medium and MMP-2 mRNA levels in cells. In studies to address the mechanisms involved in the regulation of MMP-2 activity, we found that the tissue inhibitor of matrix metalloproteinase-2 (TIMP-2), the inhibitor of MMP-2, decreased significantly when cells were treated with CTGF. Further studies showed that extracellular signal-regulated kinase (ERK) signaling is responsible for most of the CTGF-induced MMP-2 expression and TIMP-2 suppression. When NRK-49F fibroblasts were incubated with CTGF, activation of ERK1/2 signaling was observed. Suppression of ERK1/2 activation with nontoxic concentrations of PD98059, a specific inhibitor of ERK activation, was associated with a reduction of CTGF-stimulated MMP-2 activity and protein expression. In addition, the CTGF-mediated reduction of TIMP-2 activity and protein expression was prevented when ERK1/2 activation was inhibited by PD98059. These results provide evidence that CTGF augments activation of MMP-2 through an effect on MMP-2 protein expression and TIMP-2 suppression, and that these effects are dependent on the activation of the ERK1/2 pathway.     

Expression of matrix metalloproteinase-7 and tissue inhibitor of metalloproteinase-2 in human endometrial carcinoma

<正>Objective:To investigate the expression of matrix metalloproteinase-7(MMP-7) and its tissue inhibitor (TIMP-2) in endometrial carcinoma and analyze their significance in endometrial cancer's invasion and metastasis. Methods:Endometrial tissues were collected from 64 patients with endometrial carcinoma,20 patients with endometrial hyperplasia and 20 normal women.The expressions of MMP-7,TIMP-2 in endometrium were measured by immuohistochemistry. Results;Expressions of MMP-7,TIMP-2 in endometrium of patients with endometrial carcinoma were significantly higher than those in normal endometrium(P0.05).MMP-7 expression increased with surgical-pathological staging,depth of myometrial invasion,histologic grades and lymph node metastasis(P0.05),while TIMP-2 expression was related to lymph node metastasis(P0.05).TIMP-2 expression in endometrial cancer was significantly higher than that in hyperplastic endometrium(P0.05).Expressions of TIMP-2 and MMP-7 in endometrium of patients with endometrial carcinoma were positively correlated(r=0.654,P0.001). Conclusion:Highly expressed MMP-7 and TIMP-2 in endometrium may be related to development,invasion and metastasis of endometrial cancers.     

Promotion of acellular dermal matrix resolution in vitro by matrix metalloproteinase-2

OBJECTIVE: To determine whether acellular human dermis is degraded by matrix metalloproteinases (MMPs), a large class of matrix-degrading enzymes. METHODS: The degradation of acellular human dermis specimens was evaluated in vitro. Wild-type murine fibroblasts with a broad-spectrum MMP inhibitor, GM6001, and MMP-2-deficient fibroblasts were placed on the basement membrane and dermal surfaces of acellular human dermis. Matrix degradation and fibroblast infiltration into the matrix were assessed after a 20-day incubation period. RESULTS: The basement membrane thickness of the specimens cultured with wild-type fibroblasts was significantly less than that of specimens cultured with GM6001 (P<.001), and the infiltration of fibroblasts into the dermal surface was limited by the addition of GM6001 (P=.002). To determine whether MMP-2 was involved in this in vitro phenotype, MMP-2-deficient fibroblasts were assessed in comparison with wild-type fibroblasts. Wild-type fibroblasts degraded the basement membrane surface (P<.001) and infiltrated the dermal surface (P = .003) more efficiently than did MMP-2-deficient fibroblasts. CONCLUSIONS: The results from our in vitro experiments suggest that MMPs and specifically MMP-2 may play an important role in the resorption of acellular human dermis. Addition of MMP inhibitors to implanted dermal matrices may slow fibroblast infiltration and improve their longevity in vivo.     

Mitogenic bovine whey extract modulates matrix metalloproteinase-2, -9, and tissue inhibitor of matrix metalloproteinase-2 levels in chronic leg ulcers

Matrix metalloproteinases (MMPs) and their tissue inhibitors play important roles in the wound-healing process. An imbalance in the expression of these molecules is thought to contribute to the failure of chronic ulcers to heal. We investigated whether a mitogenic bovine whey extract enriched with growth factors modulated the expression and activity of MMP-2 and -9, and the tissue inhibitor of MMP-2 (TIMP-2) in chronic leg ulcers. Wound fluids and biopsies were collected from chronic leg ulcer patients whose ulcers were treated topically for 4 weeks with placebo or mitogenic bovine whey extract at concentrations of 2.5, 10, and 20 mg/mL. The levels of MMP-2 and -9 in wound fluid samples was assessed by gelatin zymography and showed a decrease in active MMP-2 in the 2.5 and 10.0 mg/mL mitogenic bovine whey extract-treated ulcers compared with placebo (p<0.05). Immunohistochemical analysis of ulcer biopsies for MMP-2, -9, and TIMP-2 expression showed a reduction in the number of MMP-2-positive dermal fibroblasts in the mitogenic bovine whey extract-treated ulcers compared with pretreatment biopsies (p<0.05) that persisted over the course of the study. In contrast, a transient increase in the number of MMP-9- and TIMP-2-positive cells was observed in mitogenic bovine whey extract treated ulcer biopsies compared with pretreatment levels (p<0.05). These results show that topical application of mitogenic bovine whey extract was able to modulate the expression of MMP-2, -9, and TIMP-2 in chronic leg ulcers and that its constituent growth factors may have the potential to redress the proteolytic imbalance observed in nonhealing chronic ulcers.