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In these experiments, we used conditioned media generated by culturing slices of melanoma bearing lung tissues from either RNF13-KO mice or their WT littermates as chemoattractants.
Intratumoral administration of CCL21-vaults in mice bearing lung cancer enhanced leukocytic infiltrates (CXCR3+T, CCR7+T, IFNγ+T lymphocytes, DEC205+ DC), inhibited lung cancer tumor growth and reduced the frequencies of immune suppressive cells [myeloid derived suppressor cells (MDSC), T regulatory cells (Treg), IL-10 T cells].
The number of mice bearing lung foci was determined.
Contrastingly, in mice bearing lung SCC, sunitinib significantly increased tumor cell proliferation (Fig 2E).
After hypoxia exposure for 10 days, we moved the animals bearing lung cancer tumor back to normoxia.
Gene-transfected tumour cells were used to cure mice bearing lung metastases by the parental, non-transduced mammary adenocarcinoma (TSA-pc).
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However in our metastasis model, these cells are F4/80- and their recruitment is not significantly different in metastasis bearing lungs compared with normal lungs (data not shown).
At 2 and 4 days after injection, mice were sacrificed and B16F10 bearing lungs were collected.
The results showed that GM-CSF concentration was significantly decreased in RNF13-KO tumor bearing lungs.
The absence of host RNF13 lowered the concentration of GM-CSF in tumor bearing lungs, leading to enhanced invasion of melanoma cells and increased lung colonization.
Assessment of the concentration of various cytokines in tumor bearing lungs and blood did not detect significant differences between the blood of RNF13-KO and WT mice; however the levels of GM-CSF were significantly reduced in RNF13-KO tumor bearing lungs, which may have guided more B16F10 cells to migrate to the lungs.
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