γ-Glutamyl transpeptidase overexpression increases metastatic growth of B16 melanoma cells in the mouse liver

Obrador, Elena; Carretero, Julián; Ortega, Ángel; Medina, Ignacio; Rodilla, Vicente; Pellicer, José A.; Estrela, José M.

doi:10.1053/jhep.2002.30277

Cited by 85 publications

(93 citation statements)

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“…Hence, GGT expression provides tumor cells with a growth advantage at physiologic concentrations of cyst(e)ine (14). Consequently, we found that tumor GGT activity and an intertissue flow of GSH, where the liver plays a key role, regulate GSH content of B16 melanoma cells and thereby their metastatic growth (15).…”

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confidence: 79%

“…Culture of B16-F10 Melanoma Cells-Murine B16-F10 melanoma cells (from the ATCC, Manassas, VA) were cultured in serum-free Dulbecco's modified Eagle's medium (DMEM; Invitrogen), pH 7.4, supplemented with 10 mM HEPES, 40 mM NaHCO 3 , 100 units/ml penicillin, and 100 g/ml streptomycin (15). Cells were harvested by incubation for 5 min with 0.05% (w/v) trypsin (Sigma) in PBS (10 mM sodium phosphate, 4 mM KCl, 137 mM NaCl), pH 7.4, containing 0.3 mM EDTA, followed by the addition of 10% calf serum to inactivate the trypsin.…”

Section: Methodsmentioning

confidence: 99%

“…Cell numbers were determined using a Coulter Counter (Coulter Electronic Inc., Miami, FL). Cell integrity was assessed by trypan blue exclusion and leakage of lactate dehydrogenase activity (15).…”

Section: Methodsmentioning

confidence: 99%

“…One of the most important functions of GSH is to store Cys because this amino acid is extremely unstable extracellularly and rapidly auto-oxidizes to cystine (13). In rapidly growing tumors, cyst(e)ine, whose concentration in blood is low, may become limiting for GSH synthesis and cell growth (14,15). Thus, malignant cells might require alternative pathways to ensure free cyst(e)ine availability.…”

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confidence: 99%

“…Oatp1 functions as a GSH/organic solute exchanger, and MRP1 functions as an organic anion export pump, but both only account for a fraction of the total GSH released into the blood. Hepatic GSH release increases in metastatic B16 melanomabearing mice (as compared with non-tumor-bearing controls), and this increased release appears to be channeled through an Oatp1/MRP1/MRP2-independent system (15). Nevertheless, the molecular nature of this transport (21) and how metastatic cells may influence its activity are still open questions.…”

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confidence: 99%

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Intertissue Flow of Glutathione (GSH) as a Tumor Growth-promoting Mechanism

Obrador

Benlloch

Pellicer

et al. 2011

Journal of Biological Chemistry

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B16 melanoma F10 (B16-F10) cells with high glutathione (GSH) content show high metastatic activity in vivo.An intertissue flow of GSH, where the liver is the main reservoir, can increase GSH content in metastatic cells and promote their growth. We have studied here possible tumor-derived molecular signals that could activate GSH release from hepatocytes. GSH efflux increases in hepatocytes isolated from mice bearing liver or lung metastases, thus suggesting a systemic mechanism. Fractionation of serum-free conditioned medium from cultured B16-F10 cells and monoclonal antibody-induced neutralization techniques facilitated identification of interleukin (IL)-6 as a tumor-derived molecule promoting GSH efflux in hepatocytes. IL-6 activates GSH release through a methionine-sensitive/organic anion transporter polypeptide 1-and multidrug resistance protein 1-independent channel located on the sinusoidal site of hepatocytes. Specific siRNAs were used to knock down key factors in the main signaling pathways activated by IL-6, which revealed a STAT3-dependent mechanism. Our results show that IL-6 (mainly of tumor origin in B16-F10-bearing mice) may facilitate GSH release from hepatocytes and its interorgan transport to metastatic growing foci.Glutathione (L-␥-glutamyl-L-cysteinyl-glycine; GSH), 2 the most prevalent non-protein thiol in mammalian cells, is involved in many cellular functions (1, 2). GSH in cancer cells is particularly relevant in the regulation of 1) carcinogenic mechanisms; 2) sensitivity against cytotoxic drugs, ionizing radiation, and some cytokines; 3) DNA synthesis; and 4) cell proliferation and death (3,4).Early studies on the organ distribution of metastatic cells showed that less than 0.1% of circulating cells survive to cause secondary metastatic growth (5). The liver is a common site for metastasis development, and, as previously shown, a high percentage of circulating cancer cells are mechanically trapped in the liver microvasculature (6). Interaction of metastatic cancer cells with the hepatic sinusoidal endothelium and Kupffer cells activates local release of proinflammatory cytokines, which then may act as molecular signals promoting cancer cell adhesion, invasion, and proliferation (3, 7). Direct in vitro lysis of metastatic tumor cells by cytokine-activated murine vascular endothelial cells has also been shown (8).From the original subcutaneous B16 melanoma tumor, spontaneously originated in C57BL/6J mice, and using sequential in vitro-in vivo growing cycles, I. J. Fidler (9) obtained 10 different cell variants (F1-F10) with increasing metastatic potentials. The B16-F10 line showed the highest metastatic activity and became a classical model widely used in metastasis research. Recent results identified the existence of a natural defense mechanism against cancer metastasis whereby the arrest of tumor cells in the liver induces endogenous NO and H 2 O 2 release, leading to sinusoidal tumor cell killing and reduced hepatic metastasis formation (3, 10). We have shown that GSH protects circulati...

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confidence: 79%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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