In-vitro resistance of cloned human glioma cells to natural killer activity of allogeneic peripheral lymphocytes.

Cells from an established culture of a human astrocytoma were incubated with normal allogeneic peripheral lymphocytes (PBL) in order to study the natural killer (NK) sensitivity of the in vitro propagated cell line. A proportion of cells in culture formed halos, into which lymphocytes did not penetrate. These cells were successfully cloned and showed a decreased susceptibility to NK cytolysis compared with the parent line. Both cell lines could be transplanted into athymic nude mice. The cloned NK-resistant cells underwent a frequent spontaneous regression in nu/nu mice, despite the fact that when used as targets for nu/nu NK cells in vitro they were only moderately susceptible. Phase-contrast microscopy of the mass-cultured cells co-cultivated with lymphocytes suggested that their morphology and ability to form inpenetrable translucent halos might influence their susceptibility to NK lysis. Experiments performed on this assumption revealed that quiescent and halo forming tumour cells were not the primary targets for NK lysis. Cells in mass culture, although tumorigenic, were thus heterogeneous in respect of susceptibility to NK attack. These findings might be relevant to the mechanism of immune escape and tumour heterogeneity in respect of spontaneous cell-mediated lysis

Luminal and basal-like breast cancer cells show increased migration induced by hypoxia, mediated by an autocrine mechanism

<p>Abstract</p> <p>Background</p> <p>Some breast cancer patients receiving anti-angiogenic treatment show increased metastases, possibly as a result of induced hypoxia. The effect of hypoxia on tumor cell migration was assessed in selected luminal, post-EMT and basal-like breast carcinoma cell lines.</p> <p>Methods</p> <p>Migration was assessed in luminal (MCF-7), post-EMT (MDA-MB-231, MDA-MB-435S), and basal-like (MDA-MB-468) human breast carcinoma cell lines under normal and oxygen-deprived conditions, using a collagen-based assay. Cell proliferation was determined, secreted cytokine and chemokine levels were measured using flow-cytometry and a bead-based immunoassay, and the hypoxic genes HIF-1α and CA IX were assessed using PCR. The functional effect of tumor-cell conditioned medium on the migration of neutrophil granulocytes (NG) was tested.</p> <p>Results</p> <p>Hypoxia caused increased migratory activity but not proliferation in all tumor cell lines, involving the release and autocrine action of soluble mediators. Conditioned medium (CM) from hypoxic cells induced migration in normoxic cells. Hypoxia changed the profile of released inflammatory mediators according to cell type. Interleukin-8 was produced only by post-EMT and basal-like cell lines, regardless of hypoxia. MCP-1 was produced by MDA-MB-435 and -468 cells, whereas IL-6 was present only in MDA-MB-231. IL-2, TNF-α, and NGF production was stimulated by hypoxia in MCF-7 cells. CM from normoxic and hypoxic MDA-MB-231 and MDA-MB-435S cells and hypoxic MCF-7 cells, but not MDA-MB-468, induced NG migration.</p> <p>Conclusions</p> <p>Hypoxia increases migration by the autocrine action of released signal substances in selected luminal and basal-like breast carcinoma cell lines which might explain why anti-angiogenic treatment can worsen clinical outcome in some patients.</p