MilliporeSigma
  • Hypoxia-induced nitric oxide release by luminal cells stimulates proliferation and uPA secretion of myoepithelial cells in a bicellular murine mammary tumor.

Hypoxia-induced nitric oxide release by luminal cells stimulates proliferation and uPA secretion of myoepithelial cells in a bicellular murine mammary tumor.

Journal of cancer research and clinical oncology (2015-02-18)
Martin Alejandro Krasnapolski, Catalina Lodillinsky, Elisa Bal De Kier Joffé, Ana María Eiján
ABSTRACT

LM38 murine mammary adenocarcinoma model is formed by LM38-LP (myoepithelial and luminal), LM38-HP (luminal) and LM38-D2 (myoepithelial) cell lines. In a previous work, we had shown that LM38-HP and LM38-D2 cell lines are less malignant than the bicellular LM38-LP cell line. To study the role of nitric oxide (NO) as one of the mediators of functional interactions between malignant luminal and myoepithelial cells. Using immunohistochemistry, in vivo iNOS expression was only detected in the luminal cells of bicellular LM38-LP and most cells of LM38-HP tumors. In cobalt-induced pseudohypoxia, LM38-LP and LM38-HP cell lines significantly increased HIF-1α and iNOS expression (Western blotting) and therefore NO production (Griess method). This increase was inhibited by the iNOS inhibitor 1400 W. On the other side, NO was not detectable in LM38-D2 cells either in basal or in pseudohypoxia. In addition, pseudohypoxia increased urokinase-type plasminogen activator (uPA) secretion by LM38-LP and LM38-HP cells and migration in the LM38-LP cell line, without modulating these properties in LM38-D2 cells (radial caseinolysis). The NO donor DETA/NONOate (500 μM) was able to increase uPA secretion and in vitro growth of LM38-D2. In agreement, 1400 W prevented in vivo growth of the myoepithelial LM38-D2 cells. Hypoxia leads to an enhanced NO production by the luminal component, through HIF-1α and iNOS, which can stimulate myoepithelial cell proliferation and uPA secretion. In these new conditions, myoepithelial cells might act as an invasive forefront generating gaps that could help luminal cells to escape from the primary tumor.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cobalt(II) chloride, AnhydroBeads, −10 mesh, 99.995% trace metals basis
Sigma-Aldrich
Cobalt(II) chloride, AnhydroBeads, −10 mesh, 99.9% trace metals basis
Sigma-Aldrich
Trypan Blue, ≥80% (HPLC), Dye content 60 %
Sigma-Aldrich
Sulfanilic acid, ACS reagent, 99%
Sigma-Aldrich
Sulfanilic acid, puriss. p.a., ≥99.0% (T)
Sigma-Aldrich
Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Triton X-305 solution, 70% in H2O
Sigma-Aldrich
Triton X-102
Sigma-Aldrich
Trypan Blue, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
HEPES buffer solution, 1 M in H2O
Sigma-Aldrich
HEPES, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)
SAFC
HEPES
SAFC
HEPES
Sigma-Aldrich
HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
Sigma-Aldrich
Triton X-100 solution, BioUltra, for molecular biology, ~10% in H2O
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
Sodium nitrite, 99.999% trace metals basis
Sigma-Aldrich
Triton X-100, peroxide- and carbonyl-free
Sigma-Aldrich
L-Arginine, reagent grade, ≥98%
Sigma-Aldrich
Triton X-45
Sigma-Aldrich
L-Arginine, 99%, FCC, FG
Sigma-Aldrich
L-Arginine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
Triton X-100, BioXtra
SAFC
L-Arginine
Sigma-Aldrich
L-Arginine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
1400W, A selective, cell-permeable, irreversible, slow, tight-binding inhibitor of inducible nitric oxide synthase (iNOS; Kd = 7 nM).