An in vitro model demonstrates the potential of neoplastic human germ cells to influence the tumour microenvironment

Autor:	Kate L Loveland, Bruce E. Loveland, Mark P. Hedger, Britta Klein, Hans-Christian Schuppe, Martin Bergmann
Rok vydání:	2017
Předmět:	Male 0301 basic medicine Cell signaling Cell Survival Urology Endocrinology Diabetes and Metabolism medicine.medical_treatment Cell Cell Communication Biology 03 medical and health sciences 0302 clinical medicine Endocrinology Immune system Testicular Neoplasms Cell Line Tumor Tumor Microenvironment medicine Humans RNA Messenger Tumor microenvironment Interleukin-6 Intratubular germ cell neoplasia Seminoma medicine.disease Coculture Techniques Germ Cells 030104 developmental biology Cytokine medicine.anatomical_structure Reproductive Medicine Cell culture Culture Media Conditioned 030220 oncology & carcinogenesis Immunology Leukocytes Mononuclear Cancer research
Zdroj:	Andrology. 5:763-770
ISSN:	2047-2919
DOI:	10.1111/andr.12365
Popis:	Testicular germ cell tumours (TGCT) typically contain high numbers of infiltrating immune cells, yet the functional nature and consequences of interactions between GCNIS (germ cell neoplasia in situ) or seminoma cells and immune cells remain unknown. A co-culture model using the seminoma-derived TCam-2 cell line and peripheral blood mononuclear cells (PBMC, n = 7 healthy donors) was established to investigate how tumour and immune cells each contribute to the cytokine microenvironment associated with TGCT. Three different co-culture approaches were employed: direct contact during culture to simulate in situ cellular interactions occurring within seminomas (n = 9); indirect contact using well inserts to mimic GCNIS, in which a basement membrane separates the neoplastic germ cells and immune cells (n = 3); and PBMC stimulation prior to direct contact during culture to overcome the potential lack of immune cell activation (n = 3). Transcript levels for key cytokines in PBMC and TCam-2 cell fractions were determined using RT-qPCR. TCam-2 cell fractions showed an immediate increase (within 24 h) in several cytokine mRNAs after direct contact with PBMC, whereas immune cell fractions did not. The high levels of interleukin-6 (IL6) mRNA and protein associated with TCam-2 cells implicate this cytokine as important to seminoma physiology. Use of PBMCs from different donors revealed a robust, repeatable pattern of changes in TCam-2 and PBMC cytokine mRNAs, independent of potential inter-donor variation in immune cell responsiveness. This in vitro model recapitulated previous data from clinical TGCT biopsies, revealing similar cytokine expression profiles and indicating its suitability for exploring the in vivo circumstances of TGCT. Despite the limitations of using a cell line to mimic in vivo events, these results indicate how neoplastic germ cells can directly shape the surrounding tumour microenvironment, including by influencing local immune responses. IL6 production by seminoma cells may be a practical target for early diagnosis and/or treatment of TGCT.
Databáze:	OpenAIRE
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