MicroRNAs - 193 and -223 alter NET differentiation markers and shape inflammatory response in vitro

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Introduction: Future improvements in NETs therapy depend on treatments tailored to tumor cell characteristics, microenvironment and inflammatory status. We recently reported selective downregulation of microRNAs (miRs) -29 and -223 in serum of NET patients, suggesting a possible role in NET disease.

Aim(s): Functional evaluation of miRs towards NET markers in vitro.

Materials and methods: We performed in vitro transfections using either mature miR or small interfering RNA (siRNA) against miR -29, -193 or -223 in NET cell lines BON and QGP. Gene and protein expression of NET differentiation markers Synaptophysin, Chromogranin A, Neuron specific enolase and proliferation Ki67/PCNA were evaluated by qRT-PCR and immunofluorescence staining. Cell survival was assessed using Propidium Iodide in flow cytometry. We determined the consequence of miR alterations in NET for the cellular immune response by stimulating macrophage cell cultures with supernatant (SN) of un-/transfected BON cells and subsequent measurement of caspase-1 activation and mRNA levels of inflammatory mediators.

Conference: 18th Annual ENETS Concerence (2021)

Presenting Author:

Authors: Geisler L, Özdirik B, Mohr R, Knorr J, Tacke F,

Keywords: miR, NET, BON, in vitro, biomarker, inflammation,