甲状腺乳头状癌组织中MT1-MMP和MMP-2的表达及其临床意义

目的通过研究膜型基质金属蛋白酶-1(membrane type matrix metalloproteinase-1,MT1-MMP)及基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)在甲状腺乳头状癌组织及癌旁组织中的表达,探讨其表达在甲状腺乳头状癌发生发展中的意义。方法采用免疫组织化学SP法检测164例甲状腺乳头状癌及其癌旁组织标本中MT1-MMP和MMP-2的表达,分析二者的表达与甲状腺乳头状癌临床病理特征间的关系。结果在甲状腺乳头状癌癌旁组织中,MT1-MMP表达阳性率为11.0%(18/164),MMP-2表达阳性率为14.0%(23/164);在甲状腺乳头状癌组织中,MT1-MMP及MMP2的表达阳性率分别为61.6%(101/164)和67.7%(111/164);二者在癌组织中的表达阳性率均高于在癌旁组织中的表达阳性率(P0.05)。癌组织中MT1-MMP和MMP-2的表达与甲状腺乳头状癌的淋巴结转移相关(P0.05),MMP-2的表达还与甲状腺乳头状癌的包膜侵犯相关(P0.05);MT1-MMP和MMP-2在甲状腺乳头状癌组织中的表达呈正相关(r=0.256,P0.05)。结论 MT1-MMP和MMP-2可能参与了甲状腺乳头状癌的包膜侵犯和淋巴结转移,且MT1-MMP和MMP-2在甲状腺乳头状癌的进展过程中可能发挥协同作用。     

基质金属蛋白酶-13在骨性关节炎发病中的活性调控研究

目的 研究一氧化氮(NO)是否通过膜型基质金属蛋白酶-1(MT1-MMP)间接激活基质金属蛋白酶-13酶原(pro-MMP-13).方法 购买并传代人软骨肉瘤细胞(SW1353),用NO供体S-亚硝基-N-乙酰基青霉胺(SNAP),SNAP+NO清除荆氧合血红蛋白(OxyHb)和SNAP+组织金属蛋白酶抑制物-2(TI...     

基质金属蛋白酶与骨的发育和重建

基质金属蛋白酶通过启动骨吸收、重塑细胞外胶原基质等方式在骨发育和骨重建中发挥着重要的作用，并与代谢性骨病的发病密切相关。深入研究基质金属蛋白酶及其抑制因子的作用机制，有助于对骨发育和骨重建这一复杂过程的深入理解和认识。     

基质金属蛋白酶-1和膜型1基质金属蛋白酶在胆囊癌血管生成拟态中的表达及临床意义

目的探讨基质金属蛋白酶-1(MMP-1)和膜型1基质金属蛋白酶(MT1-MMP)在胆囊癌血管生成拟态中的表达及临床意义。方法收集手术病理确诊的原发性胆囊癌74例和胆囊腺瘤、慢性胆囊炎各10例的石蜡标本及相关临床资料,应用免疫组化测定上述病变组织的MMP-1、MT1-MMP表达。应用Kaplan-Meier生存比较、Cox风险模型分析与胆囊癌血管生成拟态(VM)的相关性和临床意义。结果①胆囊癌MMP-1、MT1-MMP表达明显高于胆囊腺瘤、胆囊炎(P0.000 1);VM(+)胆囊癌MT1-MMP表达明显高于VM(-)胆囊癌(P=0.003 9),而MMP-1则无差别。②MMP-1表达与VM(-)胆囊癌的Nevin分期(P=0.003 6)、分化程度(P=0.010)、肝转移(P=0.003)和淋巴结转移(P=0.002)正相关;MT1-MMP表达与VM(-)或VM(+)胆囊癌的Nevin分期(P=0.013或P=0.033)、浸润深度(P=0.045或P=0.035)、淋巴结转移(P=0.046或P=0.025)和肝转移(P=0.030或P=0.027)正相关;MMP-1、MT1-MMP表达与VM(-)胆囊癌正相关。③VM(+)胆囊癌MT1-MMP表达在Nevin S3~S5期(P=0.000 1)、侵犯浆膜(P=0.001)、淋巴结转移(P=0.000 2)和肝转移(P=0.004)同样分组条件下分别明显高于VM(-)胆囊癌;MMP-1仅在肝转移组,VM(+)胆囊癌表达显著高于VM(-)(P=0.038)。④无论在MMP-1、MT1-MMP单独表达阳性或二者均阳性,VM(+)胆囊癌患者术后5年生存期都明显短于VM(-)组(P=0.025 1或P=0.022 5或P=0.025 0)。Cox多因素分析表明,VM、浸润深度、淋巴结转移、肝转移、手术方式是影响胆囊癌患者预后的独立因素。结论胆囊癌MMP-1、MT1-MMP高表达,MT1-MMP表达与胆囊癌VM相关,MMP-1则与胆囊癌VM无关。MMP-1、MT1-MMP可作为评判胆囊癌Nevin分期、浸润深度、淋巴结或肝转移及判断预后的重要指标;而MT1-MMP还可作为判断胆囊癌否存在VM和VM胆囊癌预后的重要指标。     

通关胶囊对增生前列腺组织细胞外基质的影响及机制研究

目的 评价通关胶囊对良性前列腺增生(BPH)患者的增生前列腺细胞外基质(ECM)的影响,并探讨其作用机理.方法 选择具备手术指征的BPH病人30例,按就诊顺序随机分为通关胶囊组、非那雄胺组和安慰剂组.服药4周后,行TURP术,留取组织标本.另选正常前列腺标本6例,用免疫组化法结合图像分析系统研究正常组、安慰剂组、通关胶囊组和非那雄胺组前列腺组织纤维连接蛋白(FN)、胶原(CL)、基质金属蚩白酶2(MMP-2)、金属蛋白酶组织抑制因子2(TIMP-2)和模型基质金属蛋白酶1(MT1-MMP)的阳性表达.结果 安慰剂组前列腺组织的FN、CL和MT1-MMP的阳性表达较正常组显著增强,差异有统计学意义(P<0.01),MMP-2/TIMP-2差异无统计学意义(P>0.05);通关胶囊组、非那雄胺组与安慰剂组的前列腺组织比较,FN和CL的阳性表达较安慰剂组明显减弱(P<0.01),MT1-MMP和MMP-2/TIMP-2较安慰剂组明显增高(P<0.01).结论 通关胶囊能降低增生前列腺组织ECM成分,避免其沉积,其机理可能与其调节ECM成分及其相关调控因子在前列腺组织中的表达有关.     

联合检测泛素连接酶Cullin1和基质金属蛋白酶MT4-MMP在乳腺癌组织中的表达及临床意义

目的探究泛素连接酶Cullin1和基质金属蛋白酶MT4-MMP在不同类型乳腺癌组织中表达情况及临床意义。 方法2015年1月至8月,选择徐州市肿瘤医院80例乳腺癌组织及其对应的癌旁正常乳腺组织,应用免疫组织化学法和Western blotting法分别检测其中Cullin1和MT4-MMP的表达情况,并分析其相关性。 结果Cullin1、MT4-MMP蛋白在乳腺癌组织中的表达均高于癌旁正常乳腺组织,差异均有统计学意义（77.5% vs 28.8%,67.5% vs 17.5%,χ²=38.174、40.921,均P<0.01）;在乳腺癌组织中,Cullin1及MT4-MMP蛋白的表达呈正相关（P<0.05）。Cullin1及MT4-MMP在乳腺癌中的蛋白表达在分子分型、腋窝淋巴结是否转移及TNM分期组间差异有统计学意义（P<0.05）;在患者年龄、肿瘤大小、肿瘤部位、是否复发转移组间差异无统计学意义。 结论Cullin1及MT4-MMP可能参与了乳腺癌的形成及发展过程,且两者在此过程中有一定协同作用。在分型差、分期晚的乳腺癌中,Cullin1及MT4-MMP更高的表达状态提示其可能成为新的乳腺癌恶性程度指标。     

香豆雌酚对去卵巢大鼠成骨细胞MT1-MMP和TIMP-1基因表达的影响

目的通过观察香豆雌酚(Coumestrol)对去卵巢骨质疏松模型大鼠成骨细胞膜型基质金属蛋白酶-1(MT1-MMP)及其抑制因子TIMP-1(tissue inhibitors of matrix metalloproteinase-1)基因表达的作用,探讨香豆雌酚治疗绝经后骨质疏松的可能的分子机制。方法利用去卵巢大鼠骨质疏松(OVX)模型,观察香豆雌酚对骨密度和骨组织形态计量学参数的影响;行原位杂交检测OVX大鼠胫骨近端成骨细胞中MT1-MMP、TIMP-1mRNA表达,免疫组化检测骨组织MT1-MMP、TIMP-1蛋白表达。结果COU组大鼠(经coumestrol处理)第3~5腰椎骨密度(0.165±0.015)较OVX组(0.143±0.014)明显增加,骨小梁体积比(32.7±7.9)厚度(43.3±6.7)和数目(5.5±0.9)分别较OVX组(9.7±3.3)、(35.7±8.1)、(2.5±0.6)明显升高,COU组骨小梁间隔(165.4±27.1)较OVX组(591.1±113.8)明显减小;COU组成骨细胞MT1-MMP mRNA与蛋白质表达上调,而TIMP-1基因表达无差异。结论MT1-MMP低表达是发生绝经后骨质疏松的分子机制之一,香豆雌酚可使成骨细胞MT1-MMP基因表达上调,对TIMP-1表达无明显影响。     

纳米材料碘油混悬液栓塞后兔VX2肿瘤组织MT1-MMP表达改变的意义

目的 观察应用羟基磷灰石纳米粒子和超液化碘油混悬液栓塞后,兔VX2肿瘤组织膜型基质金属蛋白酶-1(MT1-MMP)表达变化的意义.方法 60只接种VX2肿瘤细胞的荷瘤兔随机分为3组:肿瘤组、超液化碘油组、羟基磷灰石纳米超液化碘油混悬液组.通过胃十二指肠动脉插管分别给予:生理盐水(1 ml/只)、超液化碘油(0.3 ml/kg)、纳米超液化碘油混悬液(0.3 ml/kg).术后3 d CT检查确认插管成功.两周后,应用免疫组织化学三步法(S-P)测定肿瘤组织MT1-MMP的表达定位,以及治疗干预后表达量的改变.RT-PCR检测MT1-MMP mRNA的表达差异,Western blot法分析MT1-MMP蛋白表达变化.结果 MT1-MMP在肿瘤细胞膜和肿瘤组织间质都有表达.肿瘤组与碘油组、纳米碘油组的阳性表达相比,差异有统计学意义(P＜0.05),而碘油组和纳米碘油组则无明显差异(P＞0.05).Western blot法检测各组蛋白量的表达也支持此结果.RT-PCR检测3组mRNA表达值的对比无明显差异(P＞0.05).结论 MT1-MMP主要表达于肿瘤细胞膜和间质.超液化碘油和(或)羟基磷灰石纳米粒子栓塞后,肿瘤组织及间质的MT1-MMP的表达上调,可能是造成栓塞后肿瘤转移复发率高的分子机制之一.     

膜型基质金属蛋白酶-1在浸润性肝细胞癌组织的表达及其意义

目的 探讨在浸润性肝细胞肝癌(HCC)中,膜型基质金属蛋白酶-1(MT1-MMP)的表达和意义.方法 随机选取2004年1月至2006年12月间根治性手术切除的HCC标本123例,通过对肿瘤的个数、包膜是否完整、门静脉有无癌栓、有无肝外转移等标准的划分,将其分为浸润转移组(74)及无浸润组(49).术前所有病例均采用酶联免疫吸附试验(ELISA)测定血清血管内皮生长因子(VEGF)水平;并通过免疫组织化学法(ABC),测定两组术后标本癌组织的MT1-MMP蛋白及肿瘤微血管密度(MVD)的表达;定量聚合酶链反应(PCR)测定癌组织MT1-MMP mRNA的表达.结果 浸润组术前血清VEGF:(1 33.89±68.56) μg/L;无浸润组:(100.64±81.37) μg/L,差异有统计学意义(P＜0.01).MT1-MMP主要定位表达在肝癌细胞的胞膜和间质.在浸润组中,MT1-MMP蛋白及mRNA表达量明显高于无浸润组(P＜0.05);浸润组MVD:11.24±1.49,无浸润组:8.11±2.51,两者比较浸润组MVD的形成量明显增多(P＜0.01).结论 在浸润性肝细胞肝癌中,MT1-MMP的表达明显增高,并伴随血清VEGF的表达上调、癌组织内新生肿瘤微血管的形成增多.MT1-MMP高表达可作为浸润性肝癌的判定指标,并提示临床预后较差.     

基质金属蛋白酶与骨关节疾病关系的研究进展

基质金属蛋白酶 (Ｍａｔｒｉｘｍｅｔａｌｌｏｐｒｏｔｅｉｎａｓｅ,ＭＭＰｓ)是一族具有降解几乎所有细胞外基质为主要功能的酶 ,存在于正常人体 ,参与伤口愈合、软骨基质的降解、骨吸收并与细胞外基质分解的疾病有关 ,诸如类风湿性关节炎、骨性关节炎、骨质疏松症、牙周病和癌肿等[1 ] 。近年研究发现 ,ＯＡ关节软骨细胞外基质合成与降解失衡是造成变性的重要原因之一。随着检测技术的发展 ,基质金属蛋白酶在这类疾病中的作用更加受到人们的重视 ,现已成为研究骨关节疾病发病机制的一个热点。本文就金属蛋白酶与骨关节疾病关系作一综述。1…     

Parathyroid hormone inhibits the expression of membrane-type matrix metalloproteinase-1 (MT1-MMP) in osteoblast-like MG-63 cells

Parathyroid hormone (PTH) can stimulate bone resorption by increasing the activity and the number of osteoclasts in bone tissue by several mechanisms. Recently, osteoblast-derived membrane-type matrix metalloproteinase-1 (MT1-MMP) has been implied to play an important role in the process of bone resorption by degrading bone matrix. In the present study, we observed the effects of PTH (1-34) on MT1-MMP production, and the role of the protein kinase A (PKA) and protein kinase C (PKC) pathways in the regulation of MT1-MMP in cultures of human osteoblast-like MG-63 cells. By Northern blot and Western immunoblot analysis, we found, unexpectedly, that PTH (1-34) inhibited MT1-MMP mRNA and protein expression in a dose- and time-dependent manner in MG-63 cells. The PKA antagonist H-89 blunted PTH (1-34)-mediated decreases in MT1-MMP protein synthesis. Forskolin, a PKA agonist, decreased MT1-MMP expression, which was similar to the action of PTH on MT1-MMP expression, in MG-63 cells. Staurosporine, a PKC inhibitor, also blocked the inhibition by PTH. We suggest that both the PKA and PKC pathways are involved in MT1-MMP downregulation by PTH. Furthermore, we found that PTH (1-34) induced the expression of receptor activator of nuclear factor (NF)-B ligand (RANKL) mRNA in a dose- and time-dependent manner in MG-63 cells, and this effect of PTH on RANKL mRNA expression was nearly parallel to the effects of MT1-MMP downregulation, implying a correlation between MT1-MMP and RANKL expression. Our findings suggest that the decreased MT1-MMP expression induced by PTH may be involved in RANKL signaling in osteoblasts, and may play a role in the activation of bone resorption.     

Expression of membrane-type 1 matrix metalloproteinase in coronary vessels of allotransplanted primate hearts

BACKGROUND: The mechanisms of intimal thickening in cardiac allograft vasculopathy (CAV) remain controversial after heart transplantation. Matrix metalloproteinase-2 (MMP-2) plays a crucial role in degrading extracellular matrix (ECM) during neointimal formation. Recently, it has been revealed that MMP-2 is activated by membrane-type 1 matrix metalloproteinase (MT1-MMP). This process involves tissue inhibitor of MMP-2 (TIMP-2), forming an MT1-MMP/TIMP-2/pro-MMP-2 complex. In this study, we hypothesize that these components contribute to the pathogenesis of CAV. METHODS: Heterotopic cardiac allografting was performed in randomly paired Japanese monkeys with an immunosuppressive regimen of intravenous administration of antihuman CD18 monoclonal antibody. The donor hearts were harvested at Days 22, 28, 40, 41, and 95 posttransplantation. We examined expression of MMP-2, MT1-MMP, and TIMP-2 of graft vessels using immunohistochemistry and protein level by western blot analysis. RESULTS: Pathologically, various degrees of neointimal formation were observed. In the allografts harvested at Days 22, 28, 40, and 41, MT1-MMP was expressed in the endothelial cells and smooth muscle cells (SMCs) in media of some arteries without histological change, accompanied by expression of MMP-2 and TIMP-2. In the severely thickened neointima of the allograft harvested at Day 95, MMP-2 and faint MT1-MMP were expressed in SMCs of severely thickened neointima and media; TIMP-2 expression was seen only in noncollagenous tissue of severely thickened neointima. MMP-2 protein was more intensely expressed in the allograft harvested at Day 95 than in the allograft harvest at Day 41, while TIMP-2 protein level was almost same in the 2 samples. CONCLUSION: We observed the simultaneous expression of MMP-2, MT1-MMP, and TIMP-2. Thus, ECM degradation triggered by MT1-MMP/TIMP-2/pro-MMP-2 complex could be a novel mechanism of CAV.     

Inhibition of matrix metalloproteinase-14 in osteosarcoma cells by clodronate

BACKGROUND: Bisphosphonates reduce the bone metastasis formation and angiogenesis but the exact molecular mechanisms involved are unclear. Progelatinase A (proMMP-2; 78 KDa) is activated up during the tumor spread and metastasis by a cell surface-associated matrix metalloproteinase (membrane-type matrix metalloproteinase [MT1-MMP] or MMP-14). MATERIAL AND METHODS: We evaluated the effects of a bisphosphonate (clodronate) on MT1-MMP mRNA expression and protein production, catalytic activity and proteolytic activation of proMMP-2 by cultured human MG-63 osteosarcoma cells. RESULTS: Clodronate, at therapeutically attainable noncytotoxic concentrations, dose-dependently inhibited phorbol myristic acetate (PMA)-induced proteolytic activation of proMMP-2 by human MG-63 osteosarcoma cells. Clodronate also downregulated the PMA-induced expression of MT1-MMP mRNA and protein production in human MG-63 osteosarcoma cells, as evidenced by Northern analysis and fluorescent immunohistochemistry. Furthermore, clodronate inhibited directly and dose-dependently MT1-MMP activity, and the MT1-MMP inhibition by clodronate was reduced in the presence of an increased (5 mM) Ca(2+) concentrations when compared to physiological (1 mM) Ca(2+) concentrations. CONCLUSION: We conclude that (1) the extracellular/cell-associated mechanism of bisphosphonate involves inhibition of MT1-MMP catalytic activity eventually by chelation, and that (2) intracellular mechanism involves downregulation of induced MT1-MMP mRNA and protein expression. The inhibition and downregulation of MT1-MMP by clodronate can be related to their ability to reduce MG-63 osteosarcoma cell invasion and spread. These findings may, at least in part, explain at molecular level the antitumor and antibone resorption activities of clodronate observed in clinical studies.     

Neutrophil-derived serine proteinases enhance membrane type-1 matrix metalloproteinase-dependent tumor cell invasion

BACKGROUND: Matrix metalloproteinase-2 degrades a variety of basement membrane components and is essential for tumor invasion. We have previously reported that membrane type-1 matrix metalloproteinase (MT1-MMP) cooperates with neutrophil-derived serine proteinases (NDPs; elastase, cathepsin G, protease-3) to activate matrix metalloproteinase-2. We therefore hypothesized that NDPs enhance tumor-cell invasion. METHODS: Clones of human HT1080 fibrosarcoma cells transfected with MT1-MMP sense (HT-SE) or antisense CDNA (HT-AS) were used. These cells express either high (HT-SE) or extremely low levels (HT-AS) of MT1-MMP relative to nontransfected HT1080 cells (HT-WT). The cells were incubated in the presence or absence of purified NDP, with or without alpha 1-antitrypsin or the MMP inhibitor batimastat. Cell invasion was measured with the use of Boyden chambers with polycarbonate membranes coated with a reconstituted extracellular matrix. RESULTS: Under control conditions HT-WT and HT-SE cells were 4-fold more invasive than HT-AS cells. The addition of NDP increased HT-WT and HT-SE cell invasion 60% to 100% but had no effect on HT-AS cells. alpha 1-antitrypsin or batimastat did not decrease the baseline invasiveness of HT-WT and HT-SE cells; however, they abrogated the stimulatory effect of NDP. CONCLUSIONS: HT1080 cell invasion depends on MT1-MMP expression. MT1-MMP overexpression does not increase invasiveness by itself. NDPs increase invasion by MT1-MMP expressing cells by activating matrix metalloproteinase-2.     

Role of membrane type 1-matrix metalloproteinase and gelatinase A in head and neck squamous cell carcinoma invasion in vitro.

The proteolytic activity of gelatinase A, a member of the matrix metalloproteinase (MMP) family, is considered to be a critical factor in tumor cell penetration of the extracellular matrix. To express catalytic activity, however, gelatinase A requires activation by another MMP, membrane type 1-matrix metalloproteinase (MT1-MMP). The head and neck squamous cell carcinoma cell line, UM-SCC-1, forms a quiescent monolayer atop collagen unless stimulated with epidermal growth factor (EGF; 3.5 nmol/L), which induces single cell invasion within 48 hours. To determine the role of the MT1-MMP/gelatinase A protease system in an in vitro stromal invasion model, expression vectors for MT1-MMP and gelatinase A were transfected into UM-SCC-1 (SCC-1/MT and SCC-1/gelA, respectively). SCC-1/MT tumor cells were found to invade in the absence of growth factor stimulation. Additionally, these cells displayed shorter onset to invasion and penetrated deeper into the collagen gel with EGF stimulation than did control vector transfectants. SCC-1/gelA cells similarly demonstrated invasion in the absence of EGF and a heightened invasive potential under EGF-stimulated conditions. These results suggest that the MT1-MMP/gelatinase A protease system participates in squamous cell carcinoma invasion of collagenous matrices.