基质金属蛋白酶M MP-2和MM P-9在子宫颈癌中表达的免疫组化研究

目的　观察基质金属蛋白酶 MMP- 2和 MMP- 9在子宫颈癌组织中的表达 ,探讨其与子宫颈癌浸润及转移的关系。方法　采用免疫组化方法对 30例子宫颈鳞癌 ,2 9例子宫颈上皮内瘤样病变 (CIN)和 16例正常子宫颈组织进行标记及分析。结果　 MMP- 2、MMP- 9在子宫颈鳞癌、CIN、正常宫颈组织细胞浆中的阳性表达分别为2 3/ 30 (76 .7% )、17/ 30 (5 8.9% ) ;13/ 2 9(4 4.8% )、18/ 2 9(6 2 .1% ) ;4/ 16 (2 5 .0 % )、4/ 16 (2 5 .0 % )。 MMP- 2鳞癌组与CIN组及正常组相比差异有极显著性 (P<0 .0 1)。 MMP- 9正常组与鳞癌组及 CIN组相比差异有显著性 (P<0 .0 5 )。MMP- 2在各组间质的基质细胞中表达均为阳性。MMP- 9在正常宫颈组织、CIN及鳞癌的基质细胞中阳性表达分别为 0 / 16 (0 % ) ;4/ 2 9(13.8% ) ;7/ 30 (2 3.3% )。各组间相比差异无显著性。MMP- 2和 MMP- 9在鳞癌细胞浆中的阳性表达与肿瘤大小、临床分期和病理分化程度均无明显相关性。结论　 MMP- 2和 MMP- 9在子宫颈鳞癌组织中的表达与正常宫颈组织相比差异均有显著性。MMP- 2和 MMP- 9的表达水平的上调可能是子宫颈鳞癌侵袭的一个早期标志 ,但不能单独作为预测子宫颈癌结局的一个标志。     

多发性骨髓瘤患者MMP-9、MMP-2表达的研究

背景与目的:近年来的研究发现基质金属蛋白酶(MMPs)与骨重建、骨质重吸收和肿瘤浸润转移等过程有关。我们检测了多发性骨髓瘤(MM)患者的MMP-9和MMP-2水平,以探讨其与MM发病的关系和临床意义。方法:用酶联免疫吸附试验(ELISA)检测了30例MM患者单个核细胞培养液的MMP-9水平、24例MM患者骨髓基质细胞(BMSCs)培养液的MMP-2水平和12例对照组MMP-9、MMP-2水平。结果:19/30例进展期MM组的MMP-9表达水平显著高于11/30稳定期MM组和12例对照组(P<0.05);14/24例进展期MM组BMSCs的MMP-2表达水平显著高于10/24例稳定期MM组和对照组(P<0.05),稳定期MM组和对照组的MMP-9、MMP-2水平无显著差别(P>0.05);12例IgG型MM患者的MMP-9和血免疫球蛋白(M蛋白)水平呈正相关(r=0.87,P=0)。rIL-6(终浓度100ng/m l)、rIL-1β(终浓度10ng/m l)并不能显著改变BMSCs分泌的MMP-2,但9例进展期MM组BMSCs与人骨髓瘤细胞系U266细胞(终浓度2×105/mL)混合培养48小时后,MMP-2水平明显增高,显著高于上述进展期和稳定期MM患者和对照组的BMCSs(均为P<0.05)。结论:MMP-9、MMP-2水平升高与MM的病情进展有关;MMP-9可反映MM患者的肿瘤负荷,可能与MM的肿瘤浸润病理过程有关;而这些过程需骨髓瘤细胞和BMSCs相互作用而完成。     

Correlation between expression of matrix metalloproteinase-2, matrix metalloproteinase-9 and angiogenesis in gastric cancer

Objective： To investigate the relationship between matrix metalloproteinase-2 （MMP-2）, MMP-9 as well as microvessel density （MVD） and their clinicopathological features in gastric cancer. Methods： The expression of MMP-2, MMP-9 and MVD were detected by immunohistochemistry SP method on 65 cases of gastric cancer tissue and 32 adjacent gastric mucosa. Results： The positive expression of MMP-2, MMP-9 and MVD in cancer group were significantly higher than those of adjacent mucosa group （P〈0.01）. The positive expression of MM P-2 and MMP-9 had closely correlation with clinical features of lymphatic metastasis, pathological type and stages （P〈0.05）, and the high level of CD 34 had correlation with metastasis and stages （P〈0.01）. The positive expression of MMP-2 and MMP-9 showed significant correlation with the level of MVD. Prognosis was mostly affected by lymphatic metastasis and stages of clinical features. The low cum survival rate was showed in the groups of positive expression of MMP-2, MMP-9 and high level of MVD. Conclusion： MMP-2, MMP-9 and MVD play an important role in metastases, invasion and prognosis of gastric cancer, which is a valuable maker to evaluate the prognosis.     

塔斯品碱对A549细胞MMP-2与MMP-9表达的影响

目的:探讨塔斯品碱对A549细胞基质金属蛋白酶MMP-2、-9表达的影响,阐明塔斯品碱抑制肿瘤血管生成的作用机制。方法:采用免疫细胞化学SABC法和RT-PCR法检测MMP-2和MMP9在A549细胞中的表达。结果:塔斯品碱对A549细胞中MMP-2、-9蛋白及其mRNA的表达均显示一定的抑制作用并有剂量依赖关系(P<0.05)。结论:塔斯品碱抑制A549细胞中与血管生成密切相关的基质金属蛋白酶MMP-2、-9的表达,有可能用于肿瘤抗血管治疗。     

Microtubule-dependent matrix metalloproteinase-2/matrix metalloproteinase-9 exocytosis: prerequisite in human melanoma cell invasion

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that cleave and degrade a wide spectrum of extracellular matrix components. By enhancing turnover of extracellular matrix, MMP activity is also known to play a key role in tumor cell invasion. Because extracellular protease activity requires efficient release of these proteases to the cellular surface, we investigated storage, transport, and exocytosis of MMP-2 and MMP-9 in human melanoma cells using immunofluorescence, electrical, and biochemical techniques. Immunolabeling of melanoma cells with antibodies specific for MMP-2 and MMP-9 led to the identification of two distinct populations of small cytoplasmatic vesicles containing MMP-2 or MMP-9, respectively. In combination with alpha-tubulin-specific antibodies, both vesicle populations were found to be aligned along the microtubular network. Moreover, the molecular motor protein kinesin is shown to be localized on most of these vesicles, providing evidence that the identified vesicles are actively propelled along microtubules toward the plasma membrane. The functional relevance of these findings is demonstrated using low dosage (5.9 nmol/L) of paclitaxel to affect the microtubular function of melanoma cells. Although cell proliferation is not altered, paclitaxel treatment impairs secretion of MMP-2/MMP-9 and significantly reduces invasive activity in our new cell invasion assay. In conclusion, we demonstrate in melanoma cells that microtubule-dependent traffic of MMP-containing vesicles and exocytosis are critical steps for invasive behavior and therefore are potential targets for specific antitumor drugs.     

Expression and clinical significance of matrix metalloproteinase-2 and matrix metalloproteinase-9 in oral squamous cell carcinoma

To successfully establish a metastasis from an invasive carcinoma, the first step involves the degradation of the underlying basement membrane, which is mainly made up of type IV collagen. Matrix metalloproteinases (MMPs)-2 and -9 are thought to play an important role in its degradation because of their ability to destroy this type of collagen. In order to evaluate the prognostic significance of these proteases, we studied the expression of MMP-2 and -9 in series of 68 OSCC by immunohistochemistry. Of the oral carcinomas, 28% (n = 19) expressed MMP-2, and 17.6% (n = 12) expressed MMP-9. MMP-2 immunoreactivity was significantly higher in patients with alcohol consumption (p = 0.028) (OR = 4), and in those younger than 60 years (p = 0.041). MMP-9 immunostaining showed statistically significant association with the tumor grade of differentiation (p = 0.019), the T-stage (p = 0.05), and also with the alcohol intake (p = 0.04) (OR = 7.67). In the present study, although not statistically significant, we observed that immunoexpression of MMP-2 and MMP-9 was stronger in patients with lymph node metastasis (OR = 1.65 and 2.29, respectively). In patients without regional lymph node metastasis, positive MMP-9 immunostaining was related to poor survival rates (p = 0.02; OR = 5.8). MMP-2 and -9 are involved in the invasion process of oral cancer, and MMP-9 is related to poor prognosis in the subset of patients without neck node metastasis. Ethanol could enhance the carcinogenetic process in oral cavity through its influence in the expression of MMP-2 and -9.     

Relationship between expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 and invasion ability of cervical cancer cells

Constitutive overexpression of matrix metalloproteinases (MMPs) is frequently observed in malignant tumors. MMPs are a family of zinc endopeptidases consisting of at least 20 different members. In particular, MMP-2 and MMP-9 are reported to be closely associated with invasion and metastasis in several cancers. We investigated whether expression of MMP-2 and MMP-9 is associated with invasion ability of seven cervical cancer cells by administration of o-phenanthroline as MMP inhibitor. In two cell lines, Siha and Caski, MMP-2 mRNA and protein were expressed at high levels. After treatment with o-phenanthroline, the rate of invasion in these two cell lines was significantly decreased. In contrast, in the other two cell lines, HT-3 and Caski, high levels of MMP-9 mRNA and protein were expressed but there was no decrease in the rate of invasion in these cells after treatment with o-phenanthroline. The data suggest that expression level of MMP-2 mRNA may regulate with invasion ability of cervical cancer.     

基质溶解素2(MMP-10)在子宫内膜腺癌组织中的表达研究

目的:了解基质溶解素2(MMP-10)在子宫内膜腺癌组织中的表达情况,及与病变的浸润程度、组织学分级、临床分期等的关系,探讨MMP-10在子宫内膜腺癌的发病、浸润和转移中的作用及意义.方法:采用免疫组织化学方法分别检测42例子宫内膜腺癌、12例非典型增生、12例正常子宫内膜组织中MMP-10的表达,并进行统计学分析.结果:子宫内膜腺癌、子宫内膜不典型增生、正常子宫内膜组织中MMP-10阳性表达率分别为76.2%(32/42)、33.3%(4/12)、0%(0/12).MMP-10的表达呈逐渐下降趋势,每两组的统计学检验显示MMP-10在子宫内膜腺癌与子宫内膜不典型增生、子宫内膜腺癌与正常子宫内膜组织之间,均有显著性差异(P＜0.01);MMP-10的表达与组织学分级、肌层浸润程度、临床分期均有关(P＜0.01).结论:MMP-10有可能作为子宫内膜腺癌的肿瘤标志物之一.     

基质溶解素2（MMP-10）在子宫内膜腺癌组织中的表达研究

目的：了解基质溶解素2（MMP-10）在子宫内膜腺癌组织中的表达情况，及与病变的浸润程度、组织学分级、临床分期等的关系，探讨MMP-10在子宫内膜腺癌的发病、浸润和转移中的作用及意义。方法：采用免疫组织化学方法分别检测42例子宫内膜腺癌、12例非典型增生、12例正常子宫内膜组织中MMP-10的表达，并进行统计学分析。结果：子宫内膜腺癌、子宫内膜不典型增生、正常子宫内膜组织中MMP-10阳性表达率分别为76．2％（32／42）、33．3％（4／12）、0％（0／12）。MMP-10的表达呈逐渐下降趋势，每两组的统计学检验显示MMP-10在子宫内膜腺癌与子宫内膜不典型增生、子宫内膜腺癌与正常子宫内膜组织之间，均有显著性差异（P〈0．01）；MMP-10的表达与组织学分级、肌层浸润程度、临床分期均有关（P〈0．01）。结论：MMP-10有可能作为子宫内膜腺癌的肿瘤标志物之一。     

Plasma matrix metalloproteinase-9 level is better than serum matrix metalloproteinase-9 level to predict gastric cancer evolution.

PURPOSE: Matrix metalloproteinase-9 (MMP-9) in blood is a promising new tumor marker. The aims of the present study are to compare the usefulness of plasma and serum MMP-9 levels for predicting gastric cancer development, invasion, and survival. EXPERIMENTAL DESIGN: In this nested case-control study, 114 gastric cancer patients and 87 healthy controls were enrolled. MMP-9 levels and activities were quantitatively measured by ELISA assay and zymography. The results were compared with the occurrence, clinicopathologic features, and outcomes of gastric cancer patients. The follow-up time for all patients was at least 5 years. RESULTS: Serum MMP-9 levels were significantly higher than plasma MMP-9 levels. Both plasma and serum MMP-9 levels correlated significantly with active MMP-9 identified by zymography (P = 0.002 and P = 0.048, respectively). Plasma MMP-9 level was significantly elevated in gastric cancer patients when compared with control subjects (P < 0.001). Serum MMP-9 levels did not differ between the groups. Receiver-operator characteristics analysis showed the values of sensitivity (82.5%) and specificity (65.5%) at the maximum accuracy for plasma MMP-9 at >or=60 ng/mL (P < 0.001). Elevated plasma MMP-9 correlated significantly with lymph node metastasis [odds ratio (OR), 3.43; P = 0.019], lymphatic invasion (OR, 7.58; P = 0.009), and venous invasion (OR, 4.14; P = 0.033). Patients with elevated plasma MMP-9 levels had poorer survival rates than those with normal plasma MMP-9 levels (P = 0.038). Serum MMP-9 level did not correlate well with gastric cancer-invasive phenotypes or survival. CONCLUSION: Our results suggest plasma MMP-9 level is a better marker than serum MMP-9 level for predicting gastric cancer development and progression.     

Serum matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-1 in patients with malignant melanoma

Degradation of basement membranes and extracellular matrix is an essential step in cancer invasion and metastasis. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play key roles in this step. The present study was conducted to investigate the levels of MMP-3 and TIMP-1 in serum of patients with malignant melanoma and the relationship to tumor progression and known prognostic parameters. Seventy patients with cutaneous malignant melanoma were investigated. Serum samples were obtained on first admission before any adjuvant and metastatic treatment was given or follow-up of patients. Serum TIMP-1 and MMP-3 levels were determined by the solid-phase sandwich ELISA (Oncogene Science Inc.) method. The elevation of serum MMP-3 and TIMP-1 levels between the patients with malignant melanoma and healthy controls were not significantly different (p>0.05). The serum levels of MMP-3 were significantly different in males and females (p=0.001) and serum TIMP levels were influenced by age (p=0.047). Except for the ulceration status of the tumor, serum levels of MMP-3 and TIMP-1 were not related to the known prognostic factors such as tumor histology, localization, stage of the disease, Breslow thickness, Clark invasion, mitosis, TIL, and regression of tumor (p>0.05). In patients with ulceration positive, the serum levels of MMP-3 were higher (p=0.04) and TIMP-1 were lower (p=0.008) than those in patients without ulceration. No significant relationship was found between serum levels of MMP-3 and TIMP-1. In conclusion, these results suggest that neither of the serum levels of MMP-3 and TIMP-1 could be a good indicator of invasion and metastasis nor can be recommended as a tumor marker in the management of melanoma patients owing to lack of sensitivity and specificity. However, much research still continues in this field and exciting new knowledge will ultimately emerge.     

基质金属蛋白酶9和14在子宫内膜异位症中的表达

 目的　明确MMP 9和MMP 14在子宫内膜异位症中的表达。方法　利用原位杂交法检测MMP 9和MMP 14在正常子宫内膜和子宫内膜异位症病灶中的表达情况。结果　 (1)MMP 14在正常内膜中无表达 ,在异位病灶中阳性表达率为 2 0 .0 % (P <0 .0 5 ) ;(2 )MMP 9在正常内膜中仅表达于增殖期内膜 ,阳性表达率为 6 .9% ,在异位病灶中阳性表达率为 4 3.3% (P <0 .0 1)。结论　 (1)在子宫内膜异位病灶中MMP 14表达率低而MMP 9则高表达 ;(2 )MMP 14和MMP 9与内膜异位症的发生有密切关系 ,但二者间无相关性。     

Expression of RECK and matrix metalloproteinase-2 in ameloblastoma

Background  

Ameloblastoma is a frequent odontogenic benign tumor characterized by local invasiveness, high risk of recurrence and occasional metastasis and malignant transformation. Matrix metalloproteinase-2 (MMP-2) promotes tumor invasion and progression by destroying the extracellular matrix (ECM) and basement membrane. For this proteolytic activity, the endogenous inhibitor is reversion-inducing cysteine rich protein with Kazal motifs (RECK). The aim of this study was to characterize the relationship between RECK and MMP-2 expression and the clinical manifestation of ameloblastoma.     

High serum levels of matrix metalloproteinase-9 and matrix metalloproteinase-1 are associated with rapid progression in patients with metastatic melanoma.

PURPOSE: Matrix metalloproteinases (MMP) are proteolytic enzymes that play an important role in various aspects of cancer progression. In the present work, we have studied the prognostic significance of serum levels of gelatinase B (MMP-9), collagenase-1 (MMP-1), and collagenase-3 (MMP-13) in patients with advanced melanoma. EXPERIMENTAL DESIGN: Total pretreatment serum levels of MMP-9 in 71 patients and MMP-1 and MMP-13 in 48 patients were determined by an assay system based on ELISA. Total MMP levels were also assessed in eight healthy controls. The active and latent forms of MMPs were defined by using Western blot analysis and gelatin zymography. RESULTS: Patients with high serum levels of MMP-9 (> or = 376.6 ng/mL; n = 19) had significantly poorer overall survival (OS) than patients with lower serum MMP-9 levels (n = 52; median OS, 29.1 versus 45.2 months; P = 0.033). High MMP-9 levels were also associated with visceral or bone metastasis (P = 0.027), elevated serum alkaline phosphatase level (P = 0.0009), and presence of liver metastases (P = 0.032). Serum levels of MMP-1 and MMP-13 did not correlate with OS. MMP-1 and MMP-9 were found mainly in latent forms in serum, whereas the majority of MMP-13 in serum was active (48 kDa) form. MMP-13 was found more often in active form in patients (mean, 99% of the total MMP-13 level) than in controls (mean, 84% of the total MMP-13 level; P < 0.0001). After initiating the therapy, patients with elevated levels of MMP-1 (> or = 29.8 ng/mL, n = 10) progressed more rapidly than patients with lower levels (median, 1.9 versus 3.5 months; P = 0.023). Serum levels of MMP-9 and MMP-13 did not correlate with the time to progression (TTP). In multivariate analysis with age and gender, MMP-9 or MMP-1 turned out to be independent prognostic factors for OS [P = 0.039; hazard ratio (HR), 1.8; 95% confidence interval (95% CI), 1.03-3.3] or TTP (P = 0.023; HR, 2.7; 95% CI, 1.15-6.4), respectively. CONCLUSIONS: Our findings provide evidence that MMP-1, MMP-9, and MMP-13 play important roles at different phases of metastatic melanoma spread and that serum MMP-9, in particular, could have clinical value in identifying patients at high risk for melanoma progression.     

胰腺癌MMP-2和TIMP-2表达与预后的关系

目的:探讨胰腺癌组织基质质属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制剂-2(TIMP-2)表达与肿瘤侵袭转移和预后的关系.方法:通过免疫组化方法检测35例胰腺癌病理标本和10正常胰腺标本MMP-2和TIMP-2的半定量表达情况及其与肿瘤临床病理参数和预后的关系,生存分析采用Kaplan-meier方法.结果:胰腺癌MMP-2和TIMP-2表达阳性率明显高于正常对照组,与淋巴结转移和临床分级密切相关,但与胰腺癌的分化程度无关,Kaplan-meier分析发现MMP-2和TIMP-2高表达患者的生存时间明显低于MMP 2和TIMP 2低表达患者(P<0.001).结论:胰腺癌MMP-2和TIMP-2表达与肿瘤侵袭转移和预后密切相关,有助于确定术后治疗方案.     

Vascular tube formation on matrix metalloproteinase-1-damaged collagen

Connective tissue damage and angiogenesis are both important features of tumour growth and invasion. Here, we show that endothelial cells maintained on a three-dimensional lattice of intact polymerised collagen formed a monolayer of cells with a cobblestone morphology. When the collagen was exposed to organ culture fluid from human basal cell tumours of the skin (containing a high level of active matrix metalloproteinase-1 (MMP-1)), degradation of the collagen matrix occurred. The major degradation products were the $3 over 4$- and $1 over 4$-sized fragments known to result from the action of MMP-1 on type I collagen. When endothelial cells were maintained on the partially degraded collagen, the cells organised into a network of vascular tubes. Pretreatment of the organ culture fluid with either tissue inhibitor of metalloproteinase-1 (TIMP-1) or neutralising antibody to MMP-1 prevented degradation of the collagen lattice and concomitantly inhibited endothelial cell organisation into the vascular network. Purified (activated) MMP-1 duplicated the effects of skin organ culture fluid, but other enzymes including MMP-9 (gelatinase B), elastase or trypsin failed to produce measurable fragments from intact collagen and also failed to promote vascular tube formation. Together, these studies suggest that damage to the collagenous matrix is itself an important inducer of new vessel formation.