scholarly journals A rapid quantitative assay for the detection of mammalian heparanase activity

1997 ◽  
Vol 325 (1) ◽  
pp. 229-237 ◽  
Author(s):  
Craig FREEMAN ◽  
Christopher R. PARISH
Keyword(s):  
Extracellular Matrix ◽  
Cell Lines ◽  
Degradation Products ◽  
Human Cancer ◽  
Umbilical Vein ◽  
Heparan Sulphate ◽  
Metastatic Potential ◽  
Mammary Adenocarcinoma ◽  
Quantitative Assay

Heparan sulphate (HS) is an important component of the extracellular matrix and the vasculature basal laminar which functions as a barrier to the extravasation of metastatic and inflammatory cells. Cleavage of HS by endoglycosidase or heparanase activity produced by invading cells may assist in the disassembly of the extracellular matrix and basal laminar, and thereby facilitate cell migration. Heparanase activity has previously been shown to be related to the metastatic potential of murine and human melanoma cell lines [Nakajima, Irimura and Nicolson (1988) J. Cell. Biochem. 36, 157–167]. To determine heparanase activity, porcine mucosal HS was partially de-N-acetylated and re-N-acetylated with [3H]acetic anhydride to yield a radiolabelled substrate. This procedure prevented the masking of, or possible formation of, new heparanase-sensitive cleavage sites as has been observed with previous methods of radiolabelling. Heparanase activity in a variety of tissues and cell homogenates including human platelets, colonic carcinoma cells, umbilical vein endothelial cells and rat mammary adenocarcinoma cells (both metastatic and non-metastatic variants) and liver homogenates all degraded the substrate in a stepwise fashion from 18.5 to approximately 13, 8 and finally to 4.5 kDa fragments, as assessed by gel-filtration analysis, confirming the substrate as suitable for the detection of heparanase activity present in a variety of cells and tissues. A rapid quantitative assay was developed with the HS substrate using a novel method for separating degradation products from the substrate by taking advantage of the decreased affinity of the heparanase-cleaved products for the HS-binding plasma protein chicken histidine-rich glycoprotein (cHRG). Incubation mixtures were applied to cHRG–Sepharose columns, with unbound material corresponding to heparanase-degradation products. Heparanase activity was determined for a variety of human, rat and murine cell and tissue homogenates. The highly metastatic rat mammary adenocarcinoma and murine lung carcinoma cell lines had four to ten times the heparanase activity of non-metastatic variants, confirming the correlation of heparanase activity with metastatic potential. Human cancer patients had twice the serum heparanase levels of normal healthy adults. The assay will be valuable for the determination of heparanase activity from a variety of tissue and cell sources, as a diagnostic tool for the determination of heparanase potential, and for the development of specific inhibitors of heparanase activity and metastasis.

scholarly journals Biological Effects of Glucosinolate Degradation Products from Horseradish: A Horse that Wins the Race

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 343 ◽  
Author(s):  
Marijana Popović ◽  
Ana Maravić ◽  
Vedrana Čikeš Čulić ◽  
Azra Đulović ◽  
Franko Burčul ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Cell Lines ◽  
Cancer Cell ◽  
Degradation Products ◽  
Human Cancer ◽  
Biological Activities ◽  
Biological Effects ◽  
Cancer Cell Lines ◽  
Cytotoxic Activities ◽  
Minimal Inhibitory Concentrations

Horseradish degradation products, mainly isothiocyanates (ITC) and nitriles, along with their precursors glucosinolates, were characterized by GC-MS and UHPLC-MS/MS, respectively. Volatiles from horseradish leaves and roots were isolated using microwave assisted-distillation (MAD), microwave hydrodiffusion and gravity (MHG) and hydrodistillation (HD). Allyl ITC was predominant in the leaves regardless of the isolation method while MAD, MHG, and HD of the roots resulted in different yields of allyl ITC, 2-phenylethyl ITC, and their nitriles. The antimicrobial potential of roots volatiles and their main compounds was assessed against sixteen emerging food spoilage and opportunistic pathogens. The MHG isolate was the most active, inhibiting bacteria at minimal inhibitory concentrations (MICs) from only 3.75 to 30 µg/mL, and fungi at MIC50 between <0.12 and 0.47 µg/mL. Cytotoxic activity of volatile isolates and their main compounds were tested against two human cancer cell lines using MTT assay after 72 h. The roots volatiles showed best cytotoxic activity (HD; IC50 = 2.62 μg/mL) against human lung A549 and human bladder T24 cancer cell lines (HD; IC50 = 0.57 μg/mL). Generally, 2-phenylethyl ITC, which was tested for its antimicrobial and cytotoxic activities along with two other major components allyl ITC and 3-phenylpropanenitrile, showed the best biological activities.

Abstract PO-031: MetMap: A map of metastatic potential of human cancer cell lines

2020 ◽  
Author(s):  
Xin Jin ◽  
Zelalem Demere ◽  
Karthik Nair ◽  
Ahmed Ali ◽  
Gino B. Ferraro ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Human Cancer ◽  
Metastatic Potential ◽  
Cancer Cell Lines ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines

scholarly journals GHRH antagonists reduce the invasive and metastatic potential of human cancer cell lines in vitro

2010 ◽  
Vol 293 (1) ◽  
pp. 31-40 ◽  
Author(s):  
Szabolcs Bellyei ◽  
Andrew V. Schally ◽  
Marta Zarandi ◽  
Jozsef L. Varga ◽  
Irving Vidaurre ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Human Cancer ◽  
Metastatic Potential ◽  
Cancer Cell Lines ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines ◽  
Ghrh Antagonists

scholarly journals Safety studies and viral shedding of intramuscular administration of oncolytic vaccinia virus TG6002 in healthy Beagle dogs

2020 ◽  
Author(s):  
Jérémy Béguin ◽  
Virgine Nourtier ◽  
Murielle Gantzer ◽  
Sandrine Cochin ◽  
Johann Foloppe ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Cell Lines ◽  
Human Cancer ◽  
Suicide Gene ◽  
Mammary Adenocarcinoma ◽  
Toxic Metabolite ◽  
Good Tolerability ◽  
Viral Shedding ◽  
New Therapeutic Approaches ◽  
Promising Therapeutic Approach

Abstract Background: Cancer is a leading cause of mortality for both humans and dogs. As spontaneous canine cancers appear to be relevant models of human cancers, developing new therapeutic approaches could benefit both species. Oncolytic virotherapy is a promising therapeutic approach in cancer treatment. TG6002 is a recombinant oncolytic vaccinia virus deleted in the thymidine kinase and ribonucleotide reductase genes and armed with the suicide gene FCU1 that encodes a protein which catalyses the conversion of the non-toxic 5-fluorocytosine into the toxic metabolite 5-fluorouracil. Previous studies have shown the ability of TG6002 to infect and replicate in canine tumor cell lines, and demonstrated its oncolytic potency in cell lines, xenograft models and canine mammary adenocarcinoma explants. Moreover, 5-fluorouracil synthesis has been confirmed in fresh canine mammary adenocarcinoma explants infected with TG6002 with 5-fluorocytosine. This study aims at assessing the safety profile and viral shedding after unique or repeated intramuscular injections of TG6002 in seven healthy Beagle dogs.Results: Repeated intramuscular administrations of TG6002 at the dose of 5 x 107 PFU/kg resulted in no clinical or biological adverse effects. Residual TG6002 in blood, saliva, urine and feces of treated dogs was not detected by infectious titer assay nor by qPCR, ensuring the safety of the virus in the dogs and their environment.Conclusions: These results establish the good tolerability of TG6002 in healthy dogs with undetectable viral shedding after multiple injections. This study supports the initiation of further studies in canine cancer patients to evaluate the oncolytic potential of TG6002 and provides critical data for clinical development of TG6002 as a human cancer therapy.

scholarly journals Safety studies and viral shedding of intramuscular administration of oncolytic vaccinia virus TG6002 in healthy Beagle dogs

2020 ◽  
Author(s):  
Jérémy Béguin ◽  
Virgine Nourtier ◽  
Murielle Gantzer ◽  
Sandrine Cochin ◽  
Johann Foloppe ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Cell Lines ◽  
Human Cancer ◽  
Suicide Gene ◽  
Mammary Adenocarcinoma ◽  
Toxic Metabolite ◽  
Beagle Dogs ◽  
Good Tolerability ◽  
Viral Shedding ◽  
Promising Therapeutic Approach

Abstract Background Cancer is a leading cause of mortality for both humans and dogs. As spontaneous canine cancers appear to be relevant models of human cancers, the development of new therapeutic approaches can benefit both species. Oncolytic virotherapy is a promising therapeutic approach for the treatment of cancer. TG6002 is a recombinant oncolytic vaccinia virus deleted in the thymidine kinase and ribonucleotide reductase genes and armed with the suicide gene FCU1 that encodes a protein which catalyses conversion of the non-toxic 5-fluorocytosine into the toxic metabolite 5-fluorouracil. Previous studies have shown the ability of TG6002 to infect and replicate in canine tumor cell lines and its oncolytic potency in cell lines, xenograft models and canine mammary adenocarcinoma explants. Moreover, 5-fluorouracil synthesis has been confirmed in fresh canine mammary adenocarcinoma explants infected with TG6002 with 5-fluorocytosine. This study aims to assess the safety profile and viral shedding after unique or repeated intramuscular injections of TG6002 in healthy Beagle dogs. Results The maximal tolerated dose of TG6002 by intramuscular route was 5 × 107 PFU/kg. Repeated intramuscular administrations of TG6002 at the dose of 5 × 107 PFU/kg seem feasible with no particular adverse events. No clinical or biological adverse effect was observed in the course of the study. Residual TG6002 in blood, saliva, urine and feces of treated dogs was not detected by infectious titer assay and by qPCR, ensuring the safety of the virus for the dogs and their environment. Conclusions These results establish the good tolerability of TG6002 in healthy dogs with undetectable viral shedding after multiple injections. This study supports the initiation of further studies in canine cancer patients to evaluate the oncolytic potential of TG6002 and provides critical data for clinical development of TG6002 as human cancer therapy.

scholarly journals Hyaluronate Functionalized Multi-Wall Carbon Nanotubes Loaded with Carboplatin Enhance Cytotoxicity on Human Cancer Cell Lines

Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3622
Author(s):  
César Adrián Leyva-González ◽  
Daniel Salas-Treviño ◽  
Flavio Fernando Contreras-Torres ◽  
María de Jesús Loera-Arias ◽  
Christian Alexis Gómez-Tristán ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Cell Lines ◽  
Human Cancer ◽  
Human Tumor ◽  
Public Health Problem ◽  
Mammary Adenocarcinoma ◽  
Dependent Manner ◽  
Global Public Health ◽  
Multiple Cancer

Cancer is a major global public health problem and conventional chemotherapy has several adverse effects and deficiencies. As a valuable option for chemotherapy, nanomedicine requires novel agents to increase the effects of antineoplastic drugs in multiple cancer models. Since its discovery, carbon nanotubes (CNTs) are intensively investigated for their use as carriers in drug delivery applications. This study shows the development of a nanovector generated with commercial carbon nanotubes (cCNTs) that were oxidized (oxCNTs) and chemically functionalized with hyaluronic acid (HA) and loaded with carboplatin (CPT). The nanovector, oxCNTs–HA–CPT, was used as a treatment against HeLa and MDA–MB-231 human tumor cell lines. The potential antineoplastic impact of the fabricated nanovector was evaluated in human cervical adenocarcinoma (HeLa) and mammary adenocarcinoma (MDA-MB-231). The oxCNTs–HA–CPT nanovector demonstrate to have a specific antitumor effect in vitro. The functionalization with HA allows that nanovector bio–directed towards tumor cells, while the toxicity effect is attributed mainly to CPT in a dose-dependent manner.

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