B16 Melanoma Cell Arrest in the Mouse Liver Induces Nitric Oxide Release and Sinusoidal Cytotoxicity: A Natural Hepatic Defense against Metastasis
2000
The formation of liver metastases involves interactions between
intravascular cancer cells and the hepatic microvasculature. Here we
provide evidence that the arrest of intravascular B16F1 melanoma cells
in the liver induces a rapid local release of nitric oxide (NO) that
causes apoptosis of the melanoma cells and inhibits their subsequent
development into hepatic metastases. B16F1 melanoma cells (5 × 10 5 ) labeled with fluorescent microspheres were
injected into the portal circulation of C57BL/6 mice. The production of
NO in vivo was detected by electron paramagnetic
resonance spectroscopy ex vivo using an exogenous
NO-trapping agent. A burst of NO was observed in liver samples examined
immediately after tumor cell injection. The relative electron
paramagnetic resonance signal intensity was 667 ± 143
units in mice injected with tumor cells versus
28 ± 5 units after saline injection
( P < 0.001). Two-thirds of cells
arrested in the sinusoids compared with the terminal portal venules
(TPVs). By double labeling of B16F1 cells with fluorescent microspheres
and a TdT-mediated UTP end labeling assay, we determined that the
melanoma cells underwent apoptosis from 4–24 h after arrest. The mean
rate of apoptosis was 2-fold greater in the sinusoids than in the TPVs
at 4, 8, and 24 h after injection ( P < 0.05–0.01). Apoptotic cells accounted for 15.9 ± 0.8% of tumor cells located in the sinusoids and 7.1 ± 0.9% of tumor cells in the TPVs. The NO synthase inhibitor
N G -nitro-l-arginine methyl ester completely
blocked the NO burst ( P < 0.001) and
inhibited the apoptosis of B16F1 cells in the sinusoids by 77%.
However, the rate of tumor cell apoptosis in the TPVs was not changed.
There were 5-fold more metastatic nodules in the livers of
N G -nitro-l-arginine methyl ester-treated mice
( P < 0.05). The inactive enantiomer
N G -nitro-d-arginine methyl ester had no effect
on the initial NO burst or on apoptosis of tumor cells in
vivo . Both annexin V phosphatidylserine plasma membrane
labeling and DNA end labeling of apoptotic cells were demonstrated
after a 5-min exposure (a time equivalent to the initial transient NO
induction in vivo ) of B16F1 cells to a NO donor
in vitro . These results identify the existence of a
natural defense mechanism against cancer metastasis whereby the arrest
of tumor cells in the liver induces endogenous NO release, leading to
sinusoidal tumor cell killing and reduced hepatic metastasis formation.
Keywords:
- Correction
- Source
- Cite
- Save
- Machine Reading By IdeaReader
47
References
100
Citations
NaN
KQI