Anaplastic Thyroid Cancer Cells Induce the Release of Mitochondrial Extracellular DNA Traps by Viable Neutrophils.
Leonardo CristinzianoLuca ModestinoStefania LoffredoGilda VarricchiMariantonia BraileAnne Lise FerraraAmato de PaulisAlessandro AntonelliGiancarlo MaroneMaria Rosaria GaldieroPublished in: Journal of immunology (Baltimore, Md. : 1950) (2020)
Neutrophils are key effector cells that orchestrate inflammatory responses in the tumor microenvironment. Although neutrophil extracellular DNA traps (NETs) entrap and kill pathogens, they also contribute to chronic inflammation and cancer progression. Thyroid cancer (TC) is the most frequently occurring cancer of the endocrine system, accounting for 70% of deaths due to endocrine tumors. Although anaplastic TC (ATC) is rare among TCs, it is highly lethal. We demonstrated in a recent study that tumor-infiltrating neutrophil density correlated with TC size. Moreover, TC-derived soluble mediators modulate the human neutrophil phenotype. Our study aimed to investigate the involvement of NETs in human TC. Highly purified neutrophils from healthy donors were primed in vitro with a papillary TC or ATC cell line conditioned medium (CM) or with a normal thyroid CM as control. NET release was quantified using a High-Content Imaging System. Neutrophil viability was assessed by flow cytometry. Fluorescence microscopy, flow cytometry, and PCR were performed to determine the mitochondrial origin of ATC-induced NETs. ATC CM-primed neutrophils were cocultured with ATC cells to determine the effects exerted by NETs on cell proliferation. ATC CM induce NET release, whereas papillary TC or normal thyroid CM did not. ATC CM-induced NET production occurred in a reactive oxygen species-dependent and cell death-independent manner and was associated with mitochondrial reactive oxygen species production; the NETs contained mitochondrial DNA. ATC CM-primed neutrophils promoted ATC cell proliferation in a NET-dependent manner.
Keyphrases
- flow cytometry
- induced apoptosis
- cell cycle arrest
- oxidative stress
- reactive oxygen species
- cell proliferation
- cell death
- mitochondrial dna
- single molecule
- endothelial cells
- high glucose
- diabetic rats
- pi k akt
- copy number
- endoplasmic reticulum stress
- cell free
- signaling pathway
- induced pluripotent stem cells
- squamous cell carcinoma
- gene expression
- cell cycle
- mass spectrometry
- squamous cell
- drug induced
- optical coherence tomography
- genome wide
- high throughput
- multidrug resistant
- childhood cancer
- quantum dots
- young adults