Zymographic analysis of circulating and tissue forms of colon carcinoma gelatinase A (MMP-2) and B (MMP-9) separated by mono- and two-dimensional electrophoresis

Pucci‐Minafra, Ida; Minafra, Salvatore; Rocca, Giampiero La; Barranca, Marilisa; Fontana, Simona; Alaimo, Giuseppe; Okada, Yasunori

doi:10.1016/s0945-053x(01)00146-9

Cited by 56 publications

(54 citation statements)

References 16 publications

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“…In addition, the sera samples from breast cancer patients exhibited two other lytic activities of larger molecular size, similar to those previously identified as pro-MMP-9 dimers (200 kDa) and as a complex formed between pro-MMP-9 and its physiological inhibitor TIMP-1 (116 kDa) in colon cancer sera (Pucci-Minafra et al, 2001). These forms were absent or very faint in the majority of normal sera.…”

Section: Discussionsupporting

confidence: 79%

“…Moreover, most breast cancer sera display two lytic bands of 200 and 116 kDa, respectively, absent in the majority of the control sera assayed. These two lytic forms are comparable to those previously detected in colon carcinoma sera (Pucci-Minafra et al, 2001) and identified as MMP-9 dimers (the 220 kDa) and as MMP-9/TIMP1 complex (the 116 kDa).…”

Section: Gelatin Zymographysupporting

confidence: 79%

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Zymographic detection and clinical correlations of MMP-2 and MMP-9 in breast cancer sera

et al. 2004

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Matrix metalloproteinases, in particular the gelatinases MMP-2 and MMP-9, have received great attention in recent years as putative tumour markers for clinical applications. The main reason for the observed interest is their easy detection in body fluids. Moreover, recent evidence has shown multiple functions of MMPs, rather than simply degrading ECM, which include the mobilisation of growth factors and processing of surface molecules. Several authors have reported increased levels of MMPs in a number of cancers, but clinical correlations in breast cancer are still fragmentary. Thus, the aim of the present research was to investigate the activity levels of circulating gelatinases in the sera of breast cancer patients by means of zymographic analysis, and correlate data with clinicopathological parameters. In all, 80 patients and 22 healthy volunteers were involved in this study. Sera were obtained prior to surgery. The clinical variables were: grading of tumours, tumour size, lymph node involvement, tumour staging, oestrogen and progesterone receptor levels (76 out of 80 cases), and c-erbB-2 levels (46 cases). The densitometric measures of MMP-2 and MMP-9 activity levels indicated that the average values of both gelatinase activities were significantly higher in breast cancers than in control sera (Po0.0001). In addition, our analysis showed for the first time that elevated activity levels of both gelatinases correlated only with c-erbB-2 overexpression (P ¼ 0.0273 for MMP-2 and P ¼ 0.0075 for MMP-9). An inverse correlation was observed with regard to oestrogen receptor expression (P ¼ 0.0075 for MMP-2 and P ¼ 0.0273 for MMP-9). Moreover, a borderline inverse correlation was observed between the activity levels of both enzymes and nuclear grade (P ¼ 0.0511 for MMP-2 and P ¼ 0.0794 for MMP-9). In conclusion, the present data suggest that serum measures of MMP's activity may have diagnostic value for discriminating subgroups of breast cancer patients and support the hypothesis that ERBB2 amplification and/or overexpression enhance signalling pathways that may lead to increased production of gelatinases in c-erbB-2 positive breast cancers with higher metastatic potentialities.

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Section: Discussionsupporting

confidence: 79%

Section: Gelatin Zymographysupporting

confidence: 79%

Zymographic detection and clinical correlations of MMP-2 and MMP-9 in breast cancer sera

et al. 2004

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“…Purified MMP-2 and MMP-9 [8] were used as reference. After electrophoresis, gels were washed in a washing buffer (2.5% Triton X-100, 50 mM Tris, pH 7.5) for 1 h, to remove SDS and allow a partial renaturation of the protein.…”

Section: Gelatin Zymographymentioning

confidence: 99%

“…The strips containing the focused proteins were then loaded onto 9-16% linear gradient polyacrylamide gels (SDS-PAGE) and the electrophoresis run was performed by applying a constant current of 20 mA/gel at 10°C. The protein spots were revealed by ammoniacal silver staining [8]. Silver-stained gels were analyzed with ImageMaster 2D Platinum software with the support of the ExPaSy molecular biology server.…”

Section: Two-dimensional Gel Electrophoresismentioning

confidence: 99%

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New Insights into the Occurrence of Matrix Metalloproteases -2 and -9 in a Cohort of Breast Cancer Patients and Proteomic Correlations

Cara¹,

Marabeti²,

Musso³

et al. 2018

Preprint

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Matrix metalloproteases (MMPS) are a family of well-known enzymes which operate prevalently in the extracellular domain, where they fulfil the function of remodeling the extracellular matrix. Within the about 26 family members, encoded by 24 genes in humans, MMP-2 and MMP-9, have been regarded as the primary responsibility for the basement membrane and pericellular ECM rearrangement. In cases of infiltrating carcinomas, which arise from the epithelial tissues of a gland or of an internal organ, a marked alteration of the expression and the activity levels of both MMPs is known to occur. Present investigation represents the continuation and upgrading of our previous studies, now focusing on the occurrence and intensity levels of MMP-2 and -9, and their proteomic correlations, in a cohort of 80 breast cancer surgical tissues.

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Use of plasma MMP‐2 and MMP‐9 levels as a surrogate for tumour expression in colorectal cancer patients

Tutton

George

Eccles

et al. 2003

Intl Journal of Cancer

119

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Matrix metalloproteinases, and notably the gelatinases MMP-2 and MMP-9, have important roles in tumour invasion, metastasis and angiogenesis. Our study investigates the distribution of MMP-2 and MMP-9 in colorectal cancer, the correlation with plasma levels, changes following surgical resection and whether plasma levels reflect clinical staging and disease course. MMP-2 and MMP-9 expression in 48 colorectal tumours and 13 adenomatous polyps was analysed by RT-PCR, immunohistochemistry, and quantified by ELISA of tumour lysates. Concentrations of MMP-2 and MMP-9 in plasma samples from these patients and 36 other patients who underwent curative resections were measured by ELISA prior to and 6 -12 months after surgery. MMP-2 expression was significantly increased in colorectal cancer tissues compared to matched normal colon as measured by ELISA. Active MMP-2 was localised by immunohistochemistry to regions where tumour cells invaded the muscularis with little staining in more superficial areas. Plasma MMP-2 levels were also significantly elevated in patients with colorectal cancer, with significant reductions following curative resections at all stages. Similarly, MMP-9 expression was significantly increased in colorectal cancer tissues, predominantly in the tumour stroma. Plasma levels of MMP-9 were significantly elevated at all stages in colorectal cancer patients and a significant reduction was seen following curative resections. With both MMP-2 and MMP-9, the strongest correlation with clinical staging in colorectal cancer was represented by the total plasma concentration of the enzymes, both falling to within the normal range following curative surgery. Plasma levels of these enzymes may therefore have potential as a noninvasive indicator of invasion or metastasis in colorectal cancer or as a marker of disease status during follow-up. © 2003 Wiley-Liss, Inc. colorectal cancer; metastasis; proteolysis; staging; plasma Matrix metalloproteinases are a closely related multigene family of zinc-dependent proteolytic enzymes. They have a role in normal physiological tissue remodelling and are capable of degrading all components of the extracellular matrix. There is increasing evidence that metalloproteinases play many important roles in cancer, for example, releasing and activating growth factors and other proteases and assisting tumour cells to invade the extracellular matrix. In addition, MMPs potentiate neovascularisation of tumour tissue, 1-4 with both MMP-2 and MMP-9 expression being correlated with angiogenesis in colorectal cancer. 5 Several previous studies have examined MMPs and also tissue inhibitors of MMPs (TIMPs) in colorectal cancer and in some cases a relationship between expression and tumour characteristics or metastasis has been found. 2,6 -9 Other studies have measured serum or plasma levels of MMPs and stated that they are raised in various cancers; 10 -14 however few studies have been reported in colorectal cancer, and it is not clear whether circulating MMP levels reflect tumour expression....

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Zymographic analysis of circulating and tissue forms of colon carcinoma gelatinase A (MMP-2) and B (MMP-9) separated by mono- and two-dimensional electrophoresis

Cited by 56 publications

References 16 publications

Zymographic detection and clinical correlations of MMP-2 and MMP-9 in breast cancer sera

Zymographic detection and clinical correlations of MMP-2 and MMP-9 in breast cancer sera

New Insights into the Occurrence of Matrix Metalloproteases -2 and -9 in a Cohort of Breast Cancer Patients and Proteomic Correlations

Use of plasma MMP‐2 and MMP‐9 levels as a surrogate for tumour expression in colorectal cancer patients

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