Imbalance between expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in invasiveness and metastasis of human gastric carcinoma

AIM: The expressive balance between matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor of metalloproteinase-1 (TIMP-1) plays a critical role in maintaining the degradation and synthesis of extracellular matrix. Loss of such balance is associated with invasion and metastasis of tumors. This study aimed to determine the expression of MMP-9 and TIMP-1 in gastric carcinoma, and the association of the expressive imbalance between MMP9 and TIMP-1 with the invasion and metastasis and prognosis of gastric carcinoma.METHODS: We used immunohistochemistry to determine the expressions of MMP-9, TTMP-1 and proliferating cell nuclear antigen Ki-67 in the gastric specimens taken from 256 patients with primary gastric carcinoma. The patients were followed-up for up to 96 months.RESULTS: No association between the expression of MMP9 and TIMP-1 and patients' sex and age, tumor size and location of gastric carcinoma was observed. The incidence of the positive expression of MMP-9 in cases with tumors invasion to muscularis propria and visceral peritoneum (70.13% and 69.09%, respectively) was significantly higher than that in cases with tumor invasion only to lamina propria or submucosa (42.50 %, P=0.0162). The positive correlation between MMP-9 expression and the depth of tumor invasion was observed (Pearson correlation coefficient=0.2129,P=0.016). Along with the increase of the metastatic station of lymph nodes, the incidence of the MMP-9 expression was increased by degrees; a positive correlation between them was observed (Pearson correlation coefficient=0.2910,P=0.0001). There was also a significant correlation between MMP-9 expression and the TNM stage in gastric carcinoma (Pearson correlation coefficient=0.3027, P＜0.0001). The incidence of MMP-9 expression in stage Ⅱ and Ⅲ/Ⅳ (75.00%and 76.15%, respectively) was significantly higher than those in stage Ⅰ (46.15 %, P＜0.0001). A negative correlation between TIMP-1 immunoreactivity and the depth of invasion,status of lymph node metastasis and TNM stage was observed (Pearson correlation coefficient =-0.1688, -0.3556and -0.3004, P=0.023, ＜0.0001 and ＜0.0001, respectively).Four types of co-expression of MMP-9 and TIMP-1 were observed; i.e. MMP-9 positive but T IMP-1 negative (n=115),both positive (n=52), both negative (n=62) and MMP-9negative but TIMP-1 positive (n=27). The frequency of serosal invasiveness was significant higher in patients with MMP-9 but without TIMP-1 expression than those with other types of the co-expression (P=0.0303). The incidence of lymph node metastasis was highest in patients with MMP-9but without TIMP-1 expression, and lowest in those with TIMP-1 but without MMP-9 expression (P＜0.0001). The survival rate in patients with MMP-9 but without TIMP-1expression was lower than that in those with TIMP-1 but without MMP-9 expression (P=0.0014).CONCLUSION: Our results in gastric carcinoma demonstrated a significant positive association of MMP-9 over-expression with proliferation of tumor cells, the depth of invasiveness,lymph node metastasis and TNM stage, suggesting MMP-9can serve as a molecular marker of tumor invasion and metastasis. We also demonstrate a significant negative relationship of TIMP-1 expression with the depth of invasiveness and lymph node metastasis, which provide a new idea in the tumor biological and genetic treatment.The interaction between MMP-9 and TIMP-1 in the processes of tumor invasion and metastasis is that MMP-9 mainly promotes tumor invasion and metastasis and TIMP-1 inhibits functions of MMP-9. The imbalance between MMP-9 and TIMP-1 expression may suggest the occurrence of tumor invasion and metastasis, predict poor prognosis. For patients with imbalanced MMP-9 and TIMP-1 expression, the optimal treatment scheme needs to be selected.     

Relation of Matrix Metalloproteinase-9 to Different Stages of Tumors in the Serum of Gastric Cancer

The sera of 40 patients with gastric cancer and 11 patients as the control group were studied for the detection of matrix metalloproteinases. The malignant behavior of tumor cells mainly depends on the capability of invasion and the metastasis of cancer cells. After the components of the extracellular matrix (ECM) are degraded, tumor cells invade the surrounding tissue and the vascular or lymphatic vessels to form metastasis colonies at distant sites. Extracellular proteolytic enzymes implicate the invasion and metastasis of cancer. Proteolytic enzymes from tumors lead to the breakdown of basement membranes and the ECM, thereby, facilitating cancer cell invasion into the surrounding normal tissue. Proteinase activity is overexpressed in the serum of stage 3 and 4 tumors compared to stage 1 and 2 tumors. ProMMP-9 was detected in the serum of patients of gastric cancer by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-PAGE zymography, with its molecular mass of 92 kDa. MMP-9 in serum plays an important role in the progression of gastric cancer.     

基质金属蛋白酶-9和组织金属蛋白酶抑制剂-1在食管鳞癌中的表达

目的探讨基质金属蛋白酶-9（MMP-9）和组织金属蛋白酶抑制剂-1（TIMP-1）在食管鳞癌中的表达及其临床意义。方法用免疫组化和Western blot法分别检测41例食管鳞癌患者的癌及相应正常组织中MMP-9和TIMP-1的表达变化。结果食管鳞癌组织中MMP-9阳性表达率与食管癌淋巴结及静脉转移有关；MMP-9的阳性表达率与表达量均显著高于TIMP-1；MMP-9和TIMP-1的表达呈负相关。结论MMP-9与食管鳞癌的侵袭转移有关，其机制可能与食管鳞癌组织中的MMP-9／TIMP-1平衡失调有关；MMP-9与TIMP-1联合检测有助于食管鳞癌生物学行为的判断。     

基质金属蛋白酶及其组织抑制剂与食管癌浸润转移的关系

目的:研究基质金属蛋白酶-9(MMP-9)及其相应的组织金属蛋白酶抑制剂-1(TIMP-1)在食管癌组织上的表达及其在细胞浸润转移过程中所起的重要作用。方法:采用免疫组织化学链霉素抗生物素蛋白-过氧化物酶(SP)法检测的50例食管癌组织中MMP-9及TIMP-1的表达。结果:MMP-9在食管癌组织的阳性表达率为76％,其表达与淋巴结转移呈正相关(Pearson列联系数0.343,P<0.05)。TIMP-1在食管癌组织表达的阳性率为30％,其表达与淋巴转移呈负棚关(Pearson列联系数为O.333,P<0.05)。结论:MMP-9阳性表达与食管癌浸润转移呈正相关,TIMP-1阳性表达与食管癌浸润转移呈负相关,二者在食管癌浸润转移中的关系表现为MMP-9促进肿瘤转移,TIMP-1对食管癌有独立的抑制作用。通过检测食管癌组织中的MMP-9及TIMP-1的表达,对预后的评价有一定价值。     

Peripheral blood level alterations of TIMP-1, MMP-2 and MMP-9 in patients with type 1 diabetes.

AIM: To determine the plasma levels of enzymes and inhibitors involved in extracellular matrix turnover in patients with Type 1 diabetes with normal renal function. METHODS: Plasma levels of matrix metalloproteinases 2 and 9 (MMP-2, MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) were measured in 43 Type 1 diabetic subjects and age- and sex-matched controls. RESULTS: No significant difference in plasma MMP-2 between diabetic patients and controls was observed. MMP-9 was detected in the plasma of 15 diabetic patients (35%), but undetectable in all control subjects (P < 0.015). Plasma TIMP-1 concentrations were significantly elevated (P < 0.001) in diabetic patients compared to controls. There was no correlation observed between MMP-2, MMP-9 and TIMP-1 and similarly between MMP-2, MMP-9 and TIMP-1 and age, duration of diabetes, blood pressure and glycated haemoglobin (HbA1c). CONCLUSIONS: This study has demonstrated alterations in several plasma extracellular matrix modulators in the absence of significant vascular disease.     

The expression of matrix metalloproteinases (MMPs), tissue inhibitor of metalloproteinases (TIMPs) and angiogenesis in relation to the depth of tumor invasion and lymph node metastasis in submucosally invasive colorectal carcinoma]

BACKGROUND/AIMS: Lymph node (LN) metastasis occurs in approximately 10% of patients with submucosally invasive colorectal carcinoma. This study was performed to determine the role of matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs) production and microvessel formation on the LN metastasis in submucosally invasive colorectal carcinoma. METHODS: A total of forty-one subjects with surgically resected submucosally invasive colorectal carcinoma were included in this study. Immunohistochemical staining of MMP-2, MMP-9, TIMP-1, TIMP-2, and urokinase-type plasminogen activator were performed. Angiogenesis was evaluated by counting the number of microvessels in each pathologic specimen as identified by CD34 immunohistochemical staining. RESULTS: The depth of submucosal invasion was not significantly correlated with the expression of MMP-2, MMP-9, TIMP-1, TIMP-2, or urokinase-type plasminogen activator, but the microvessel count was significantly correlated with the absolute depth of invasion (r=0.312, p<0.05). Upregulation of TIMP-2 was positively correlated with adjacent lymphatic invasion (p<0.05) and increased TIMP-2 expression was correlated with LN metastasis in submucosally invasive colorectal carcinoma (p=0.088). CONCLUSIONS: These results suggest that the expression of TIMP-2 and the microvessel count may be useful parameters for considering additional surgery after endoscopic treatment of submucosally invasive colorectal carcinoma.     

胃肠安方对胃癌基质金属蛋白酶9及基质金属蛋白酶抑制因子1/2蛋白表达的影响

[目的]观察胃肠安方对人胃癌裸鼠原位移植瘤转移的抑制作用及其对基质金属蛋白酶9 (matrix metalloproteinase-9,MMP-9)、基质金属蛋白酶抑制因子(matrix metalloproteinases tissue inhibitor,TIMP)-1、TIMP-2蛋白表达的影响.[方法]建立裸小鼠胃原位癌模型,分为胃肠安方组及模型组,观察裸小鼠胃癌种植后肿瘤转移灶情况以及与肿瘤生长与转移密切相关的MMP-9、TIMP-1、TIMP-2的影响.[结果]胃肠安方能够抑制胃癌生长与转移,与模型组比较,差异具有统计学意义(P＜0.05).胃肠安方组MMP-9蛋白阳性表达明显弱于模型组,TIMP-1及TIMP-2蛋白阳性表达明显强于模型组.[结论]胃肠安方具有抑制人胃癌裸鼠原位移植瘤转移作用,其抑制胃癌转移的部分作用机制与抑制MMP-9蛋白表达及上调TIMP-1、TIMP-2蛋白表达有关.     

胃癌组织中MMP-9和TIMP-1的表达及其临床意义

背景：有关胃癌组织中基质金属蛋白酶（MMPs）和金属蛋白酶组织抑制剂（TIMPs）表达的研究不多且结果不一。目的：探讨胃癌组织中MMP-9和TIMP-1的表达及其临床意义。方法：收集临床病理资料完整的98例胃癌患者。应用免疫组化法检测组织中MMP-9和TIMP-1表达．并分析其与临床病理特征的关系以及各参数对胃癌患者预后的影响。结果：胃癌组织中MMP-9高表达与浸润深度、淋巴结转移和TNM分期显著相关（P〈0．01）。单因素分析显示胃癌患者预后与MMP-9高表达（P=0．014）、浸润深度（P〈0．001）、淋巴结转移（P〈0．0001）和TNM分期（P〈0．0001）相关。TIMP-1表达与胃癌患者临床病理特征和预后无关。结论：MMP-9高表达与胃癌的发生、进展有关,可作为胃癌患者预后指标之一。     

The diagnostic value of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of matrix metalloproteinases 1 (TIMP-1) determination in the sera of colorectal adenoma and cancer patients

Purpose  

Tumor cells, including colorectal cancer (CRC), are able to produce and release matrix metalloproteinase 9 (MMP-9) which is involved in tumor invasion and metastasis. Natural tissue inhibitors of matrix metalloproteinases (TIMPs) regulate activity of MMPs and stimulate tumor growth and malignant transformation. The aim of the present study was to compare the clinical significance of serum MMP-9 with TIMP-1 in the diagnosis of CRC patients and in the differentiation between colorectal adenoma (CA) and cancer.     

Enhanced Cell Surface Expression of Matrix Metalloproteinases and Their Inhibitors, and Tumor-Induced Host Response in Progression of Human Gastric Carcinoma

The cell surface and/or intracellular expression of the matrix metalloproteinases (MMP -2, 7, and -9 and MT1-MMP) and their inhibitors (TIMP-2 and -4) were investigated in tumor and tumor-infiltrating lymphocytes (TIL) in gastric carcinoma (n = 15) from the primary locus, metastatic gastric carcinoma (n = 20) from malignant ascites, and benign gastric mucosa (n = 20) for the control. The quantitative analysis was based on the percentage of positive cells by flow cytometry. The results clearly showed increased cell surface expression of MMP-2, -7, and -9, MT1-MMP, and TIMP-2 and -4 in both tumor cells and TIL during the development of invasion and/or metastasis of gastric carcinoma. There were equilateral correlations with cancer progression and frequency of cell surface expression of MMPs and their inhibitors, TIMPs, suggesting not only the aggressive nature of particularly metastatic gastric carcinoma, but also the presence of MMPs complexed with TIMPs on tumor cells and TIL. The enhanced cell surface expression of MMPs and TIMPs on TIL within metastatic carcinoma nests showed the result of a host response induced by tumors. These suggest that the increased cell surface expression of MMPs and TIMPs, and tumor-induced host response play a key role in gastric cancer invasion and/or metastasis.     

Determination of the serum matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in patients with either advanced small-cell lung cancer or non-small-cell lung cancer prior to treatment

This study was undertaken to examine the relationship between circulating matrix metalloproteinase (MMP-9) and tissue inhibitor of matrix metalloproteinase (TIMP-1) in our patients with either advanced small-cell lung cancer (SCLC) or non-small-cell lung cancer (NSCLC) prior to treatment. Thirty-one male and female patients with either stage III or IV NSCLC and 17 with either stage III or IV SCLC were compared to 117 age matched non-smoking controls of both sexes. Prior to any treatment of the patient, a baseline serum sample was obtained from each of the patients for the determination of circulating MMP-9 and TIMP-1 by ELISA. The results indicate that both MMP-9 and TIMP-1 are elevated in the serum of lung cancer patients when compared to the controls. This observation was true for both SCLC and NSCLC. However, the mean values for both MMP-9 and TIMP-1 in the two tumors were not different from each other. The natural physiological relationship between MMP-9 and the inhibitor TIMP-1 was lost in both SCLC and NSCLC, indicative of abnormal alterations by the tumor. The data from this study suggests that advanced lung cancer does alter the normal circulatory pattern of MMP-9 and TIMP-1. This could aid in the processes of tumor invasion and/or metastasis.     

DNA ploidy analysis and expression of MMP-9, TIMP-2, and E-cadherin in gastric carcinoma

AIM: To investigate DNA ploidy and expression of MMP-9, TIMP-2, and E-cadherin in gastric carcinoma and to explore the mechanism of invasion and metastasis of gastric carcinoma. METHODS: Immunohistochemical methods were used to detect the expressions of MMP-9, TIMP-2, and E-cadherin in 156 cases, including 99 cases of gastric carcinoma, 16 cases of adjacent noncancerous mucosa, 16 cases of distant metastases and 25 cases of metastatic lymph node (LN) from gastric carcinoma. Flow cytometry DNA ploidy and S-phase fraction (SPF) analysis were performed on 57 cases, including 47 cases of gastric cancer, 6 cases of adjacent noncancerous mucosa, and 4 cases of distant metastatic cancer. RESULTS: The expression of MMP-9 was significantly correlated with Lauren's classification, Borrmann's classification, LN metastasis, tumor metastasis, and TNM stage, as well as depth of invasion (all P<0.05). The positive rate was lower in noncarcinoma than in carcinoma (31.3% vs66.7%, P<0.01). The expression of TIMP-2 was significantly correlated with Borrmann's classification, LN metastasis, and the depth of invasion (all P<0.05), The expression of E-cadherin was significantly correlated with differentiation, Lauren's classification, Borrmann's classification, and LN metastasis, as well as the depth of invasion (P<0,01 or P<0.05). E-cadherin was less expressed in carcinoma than in noncarcinoma (42.4% vs87.5%, P<0.01). There was a positive correlation between MMP-9 and TIMP-2 and a negative correlation between MMP-9 and E-cadherin, but no correlation between TIMP-2 and E-cadherin. Also there was a positive correlation between DNA aneuploid rate and differentiation and LN metastasis. SPF that was higher than 15% was positively correlated with tumor size, differentiation and LN metastasis. And there was a significant difference between carcinoma and noncarcinoma in DNA aneuploid rate and SPF. CONCLUSION: With tumor progression and development of heterogeneity, the abnormal expressions of MMP-9, TIMP-2, and E-cadherin or DNA aneuploid rate or high SPF gradually increases, suggesting that they play a crucial role in gastric carcinoma progression.     

Matrix metalloproteinases and their inhibitors in gastric cancer

Background—The matrix metalloproteinases (MMPs)and tissue inhibitors of matrix metalloproteinases (TIMPs) are stronglyimplicated in tumour invasion and metastasis.
Aims—To investigate the presence of individualMMPs and TIMPs in gastric cancer.
Methods—The presence of MMP-1, MMP-2, MMP-3,MMP-9, TIMP-1, and TIMP-2 was identified in a group of gastric cancers(n=74) by immunohistochemistry using monoclonal antibodies. Theseantibodies were effective on formalin fixed, paraffin wax embedded sections.
Results—A large proportion (94%) of gastriccancers contained MMP-2; MMP-1 and MMP-9 were also detected in 73% and70% of tumours respectively. MMP-3 was only present in 27% oftumours. MMP-1 and MMP-9 were found predominantly in intestinal typetumours. TIMP-1 and TIMP-2 were identified in 41% and 57% of tumoursrespectively. Immunoreactivity for individual MMPs or TIMPs was notidentified in normal stomach.
Conclusions—This study shows the presenceof matrix metalloproteinases, particularly MMP-2, and TIMPs in stomachcancer. Antibodies which are effective in formalin fixed, paraffin waxembedded sections are useful for the identification of MMPs and TIMPsin diagnostic specimens.

Keywords:immunohistochemistry; matrix metalloproteinase; neoplasm; stomach

     

Expression of Tissue Inhibitors of Metalloproteinases (TIMPs) in Gastric Cancer

Matrix metalloproteinases (MMPs) are one of the major classes of proteolytic enzymes involved in tumor invasion and metastasis, being inhibited by naturally occurring tissue inhibitors of metalloproteinases (TIMPs). Sixty-five patients who underwent surgery for gastric cancer in 1992 at Chonnam National University Hospital were selected for this study. The primary selection criteria were the availability of formalin-fixed and paraffin-embedded blocks and sufficient clinical follow-up for tumor-specific survival analysis. In this study, we examined the expression of TIMP-1 and TIMP-2 in human gastric cancer tissue by in situ hybridization and immunohistochemistry, and the correlation between their expression and clinicopathological parameters. TIMP-1 and TIMP-2 expressions were detected predominantly in the peritumor stromal cells rather than tumor cells themselves. Immunohistochemical stainings were concordant with the result obtained by in situ hybridization. The intensity of TIMP-1 immunohistochemical stromal staining correlated with tumor stage (P = 0.009) and patient survival (P = 0.025). However, the intensity of TIMP-2 immunohistochemical stromal staining did not correlate with tumor stage (P = 0.339) and patient survival (P = 0.474). The correlation between the increased TIMP-1 expression and cancer stage noted in this study reflects a role of TIMP-1 in predicting the aggressive behavior of gastric cancer. TIMP-2 expression did not correlate with clinicopathological parameters. However, expression of TIMP-1 and the possible additional value of TIMP-2 should be further explored in determining the prognosis of gastric cancer.     

Matrix metalloproteinases 2 and 9 in oral squamous cell carcinomas: manifestation and localization of their activity

Purpose The process of invasion and metastasis is closely related to the prognosis of oral squamous cell carcinoma (OSCC). Matrix metalloproteinases (MMPs) are a group of enzymes characterized by their ability to degrade extracellular matrix proteins and contribute to the tumor invasion and metastasis. Especially MMP-2 and MMP-9 are known to be related to destruction of basement membrane as collagenases. This study focused on protein expression of MMP-2 and MMP-9 and their extracellular matrix degradation activity in OSCCs.Methods Freshly frozen samples from 31 OSCC patients were analyzed for the localization and activity of MMP-2 and MMP-9. Serial frozen sections were used by routine hematoxylin and eosin staining, immunohistochemistry for MMP-2 and MMP-9, and film in situ zymography (FIZ) for gelatinolytic activity. We also evaluated the activity of MMP-2 and MMP-9 by zymography using the same samples as frozen sections. The activated form/proform ratio of MMPs in zymography was evaluated using an image scanner.Results In MMP-2 the proportion in T3 and T4 clinical stage groups was significantly higher than that in T1 and T2. The proportion in lymph node metastasis cases (N⁺) was also significantly higher than that in non-lymph node metastasis cases (N^–). In contrast to MMP-2, the activated form/proform ratio of MMP-9 was very low, suggesting that MMP-9 is not activated in the matrix degradation of OSCC, although both MMP-2 and MMP-9 protein expression are presented in tumor cells. FIZ revealed MMP in both tumor cells and stromal cells of 70% of the N⁺ cases and of 47.6% of the N^– cases.Conclusions These results indicate that two types of proform and activated form matrix metalloproteinases, MMP-2 and MMP-9, are present in human OSCC, and that the activated MMP-2 could be a main enzymatic activity of gelatinolysis in OSCC. Interaction of tumor cells and stromal cells seems to play an important role in the invasion and metastasis of human OSCC. Combination analysis of zymography and FIZ is a usuful method to detect activity and localization of MMPs in human OSCC.     

Interleukin-6 regulation of matrix metalloproteinase (MMP-2 and MMP-9) and tissue inhibitor of metalloproteinase (TIMP-1) expression in malignant non-Hodgkin's lymphomas.

We showed previously that human malignant non-Hodgkin's lymphomas (NHL) degrade extracellular matrix (ECM) components through the action of metalloproteinases and that elevated expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) correlated with a poor clinical outcome in patients with NHL. In the present study we sought to investigate whether there is any correlation between the expression of gelatinases (MMP-2 and MMP-9), TIMP-1, and the expression of cytokines and growth factors such as interleukin-1beta (IL-1beta), IL-6, IL-10, tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGFbeta), and basic fibroblast growth factor (bFGF) in human NHL. In lymphoma tissues obtained from 32 patients, elevated expression of IL-6 correlated significantly with elevated messenger RNA (mRNA) levels of MMP-9, MMP-2, and TIMP-1. Moreover, in human lymphoid cell lines of B- and T-cell origin (Raji, Jurkat, and NC-37), IL-6 stimulated production of MMP-9 and MMP-2 but not TIMP-1. In the Matrigel invasion assay IL-6 significantly upregulated transmigration of Raji and Jurkat cells, which in turn was inhibited by recombinant human TIMP-1 and anti-MMP-9 and MMP-2 antibodies. We postulate that IL-6 may play a role in the clinical aggressiveness of human NHL by stimulating MMP production.     

基质金属蛋白酶-9及其抑制物-1在原发性肝癌中的表达及临床意义

探讨肝细胞癌(HCC)中MMP-9及TIMP-1mRNA的表达,及其与HCC侵袭、转移等生物学行为的关系。使用RT-PCR法检测20例HCC及癌周组织手术切除标本的MMP-9、TIMP-1mRNA表达。结果显示HCC组的MMP-9、TIMP-1mRNA的相对含量均高于癌周对照组。有包膜浸润的HCC的MMP-9 mRNA表达明显高于无包膜浸润HCC。提示MMP-9、TIMP-1在HCC的浸润和转移中起重要作用。MMP-9 mRNA表达与肝癌细胞包膜浸润密切相关。