Assessment of Gelatinases (MMP-2 and MMP-9) by Gelatin Zymography

Tóth, Márta; Sohail, Anjum; Fridman, Rafael

doi:10.1007/978-1-61779-854-2_8

Cited by 313 publications

(280 citation statements)

References 32 publications

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“…Activity of MMP-2 was determined by zymography assay [19]. After treatment the culture medium was loaded on a SDS-polyacrylamide (10%) gel containing 1% gelatin.…”

Section: Western Blot Analysis Of Collagen and Mmp Expression And Zymmentioning

confidence: 99%

Tamoxifen induces the development of hernia in mice by activating MMP-2 and MMP-13 expression

Liu

Wang

et al. 2015

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Hernia is a disease with defects in collagen synthesis/metabolism. However, the underlying mechanisms for hernia formation have not been fully defined. Tamoxifen is a selective estrogen receptor modulator and used for patients with breast cancer. Tamoxifen also has pleiotropic and side effects. Herein, we report that tamoxifen treatment resulted in an appearance of a large bulge in the low abdomen between the hind legs in male but not in female mice. The autopsy demonstrated that the low abdominal wall was broken and a large amount of intestine herniated out of the abdominal cavity. Histological analysis indicated that tamoxifen caused structural abnormalities in the low abdominal wall which were associated with decreased type II collagen content. Furthermore, we determined increased matrix metalloproteinase-2 (MMP-2) and MMP-13 expression in the tissue. In vitro, tamoxifen induced MMP-2 and MMP-13 expression in fibroblasts. The promoter activity analysis and ChIP assay demonstrate that induction of MMP-13 expression was associated with activation of JNK-AP-1 and ERK1/2 signaling pathways while induction of MMP-2 expression was related to activation of the ERK1/2 signaling pathway. Taken together, our study establishes a novel murine hernia model, defines a severe side effect of tamoxifen, and suggests a caution to male patients receiving tamoxifen treatment.

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“…Activity of MMP-2 was determined by zymography assay [19]. After treatment the culture medium was loaded on a SDS-polyacrylamide (10%) gel containing 1% gelatin.…”

Section: Western Blot Analysis Of Collagen and Mmp Expression And Zymmentioning

confidence: 99%

Tamoxifen induces the development of hernia in mice by activating MMP-2 and MMP-13 expression

Liu

Wang

et al. 2015

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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“…The bulk hydrogels of photocrosslinked gelatin methacryloyl were used in drug delivery and tissue engineering applications for its biocompatibility (23). Gelatin also contains target cleavage sites for MMP-2, thereby appealing as a biodegradable structural material for microrobots (24). The present study also shows hydrogels as a useful material platform for microswimmers to carry concentrated therapeutics in their bulk volume.…”

Section: Introductionmentioning

confidence: 70%

3D-Printed Biodegradable Microswimmer for Drug Delivery and Targeted Cell Labeling

Ceylan

Yasa

Yaşa

et al. 2018

Preprint

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Miniaturization of interventional medical devices can leverage minimally invasive technologiesby enabling operational resolution at cellular length scales with high precision and repeatability.Untethered micron-scale mobile robots can realize this by navigating and performing in hard-toreach, confined and delicate inner body sites. However, such a complex task requires an integrated design and engineering strategy, where powering, control, environmental sensing, medical functionality and biodegradability need to be considered altogether. The present study reports a hydrogel-based, biodegradable microrobotic swimmer, which is responsive to the changes in its microenvironment for theranostic cargo delivery and release tasks. We design a double-helical magnetic microswimmer of 20 µm length, which is 3D-printed with complex geometrical and compositional features. At normal physiological concentrations, matrix metalloproteinase-2 (MMP-2) enzyme can entirely degrade the microswimmer body in 118 h to solubilized non-toxic products. The microswimmer can respond to the pathological concentrations of MMP-2 by swelling and thereby accelerating the release kinetics of the drug payload. Anti-ErbB 2 antibodytagged magnetic nanoparticles released from the degraded microswimmers serve for targeted labeling of SKBR3 breast cancer cells to realize the potential of medical imaging of local tissue sites following the therapeutic intervention. These results represent a leap forward toward clinical medical microrobots that are capable of sensing, responding to the local pathological information, and performing specific therapeutic and diagnostic tasks as orderly executed operations using their smart composite material architectures.

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“…Furthermore, most MMPs are secreted as zymogenes upon cytokine stimulation and require extracellular proteolytic activation. MMP-2 (gelatinase A, EC 3.4.24.24) and MMP-9 (gelatinase B, EC 3.4.24.35) have been extensively studied owing to their consistent association with tumor invasion and metastasis (Toth and Fridman, 2001;Verma and Hansch, 2007). In this way inhibition of expression, conversion and/or biological (enzymatic) function of these proteases may block angiogenesis both in vivo and in vitro (Kappert et al, 2009;Verma and Hansch, 2007).…”

Section: Resultsmentioning

confidence: 99%

“…It is noteworthy that 4-aryl-chromenes with methoxy substitution(s) at 4-position, have been identified as potent vascular disrupting agents (Cai et al, 2009;Kasibhatla et al, 2004). Vascular disrupting agents, which are a new class of potential anti-cancer drugs, disrupt preformed endothelial Lanes 1 and 8 represent the ConA (as positive control, note the relative induction of MMP-2 in response to ConA, at 5 lg/ml, compared to other lanes) and tetracycline (as negative control) effects on MMP activity, respectively (Toth and Fridman, 2001). Note the partial inhibition of MMP-2 (62-72 kDa) and complete suppression of MMP-9 (100 kDa), at 2 lg/ml of tetracycline (Kappert et al, 2009).…”

Section: Resultsmentioning

confidence: 99%

“…The matrix metalloproteinase-2 (MMP-2) and MMP-9 gelatinolytic activities of conditioned media from HUVECs were assayed by electrophoresis (Toth and Fridman, 2001). Confluent HUVECs were isolated and immediately incubated in the absence of FBS with different concentrations of the test compounds for 16 h. Equal protein samples concentrated serum free culture medium of HUVECs treated with 1-16 lg/ml of test compounds, measured by the method of Bradford, were separated on 7.5% SDSpolyacrylamide gels containing 2 mg/ml (0.2%) gelatin under nonreducing conditions at 4°C (Bradford, 1976).…”

Section: Gelatin Zymography Assaymentioning

confidence: 99%

See 1 more Smart Citation

Anti-angiogenic/proliferative behavior of a “4-aryl-4H-chromene” on blood vessel’s endothelial cells: A possible evidence on dual “anti-tumor” activity

et al. 2010

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Development of therapies aimed at inhibiting the angiogenesis, in combination with classical anti-cancer therapies, is among the most intensively studied approaches to the treatment of cancer. As a continuation of the efforts to discover and develop ''4-aryl-4H-chromenes'' as novel anticancer agents, in this study, we have been synthesizing (and examining) these compounds that appear to inhibit metastasis and, in particular, angiogenesis by using successful approaches such as three-dimensional capillary tube formation as well as matrix metalloproteinase (MMP) gelatinase assay in the endothelial cell-based experimental system. Anti-angiogenic and anti-proliferative effects of chromene compound 1 were especially checked on three-dimensional culture of human umbilical vein endothelial cells (HUVECs) in collagen matrix and HUVECs proliferation assay, respectively. Compound 1 was identified to be a highly potent anti-angiogenic agent at well-tolerated concentrations. In mechanistic studies, baseline MMP activities were inhibited in the presence of compound 1, in a dose-dependent manner. Based on our data, there is this possibility that chromene compounds (especially 1) are useful in treatment of some cancers because of their ability to both induce cancer cell apoptosis and reduce the stimulatory factors involved in HUVEC cell proliferation and angiogenesis.

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Assessment of Gelatinases (MMP-2 and MMP-9) by Gelatin Zymography

Cited by 313 publications

References 32 publications

Tamoxifen induces the development of hernia in mice by activating MMP-2 and MMP-13 expression

Tamoxifen induces the development of hernia in mice by activating MMP-2 and MMP-13 expression

3D-Printed Biodegradable Microswimmer for Drug Delivery and Targeted Cell Labeling

Anti-angiogenic/proliferative behavior of a “4-aryl-4H-chromene” on blood vessel’s endothelial cells: A possible evidence on dual “anti-tumor” activity

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