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Activin A target genes are differentially expressed between normal and neoplastic adult human testes: clues to gonocyte fate choice
Author(s) -
Szarek M.,
Bergmann M.,
Konrad L.,
Schuppe H.C.,
Kliesch S.,
Hedger M. P.,
Loveland K. L.
Publication year - 2019
Publication title -
andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.947
H-Index - 43
eISSN - 2047-2927
pISSN - 2047-2919
DOI - 10.1111/andr.12553
Subject(s) - seminoma , gonocyte , sertoli cell , biology , somatic cell , germ cell , testicular germ cell tumor , immunohistochemistry , rete testis , endocrinology , spermatogenesis , cell culture , medicine , cancer research , gene , immunology , genetics , epididymis , chemotherapy , sperm
Background Human testicular germ cell tumours ( TGCT ) arise from germ cell neoplasia in situ ( GCNIS ) cells that originate from foetal germ cell precursors. Activin A is central to normal foetal testis development, and its dysregulation may contribute to TGCT aetiology. Objective (i) To test whether the expression profiles of activin A targets in normal and neoplastic human testes indicates functional links with TGCT progression. (ii) To investigate whether activin A levels influence MMP activity in a neoplastic germ cell line. Materials and Methods (1) Bouin's fixed, paraffin‐embedded human testes were utilized for PCR ‐based transcript analysis and immunohistochemistry. Samples ( n = 5 per group) contained the following: (i) normal spermatogenesis, (ii) GCNIS or (iii) seminoma. CXCL 12, CCL 17, MMP 2 and MMP 9 were investigated. (2) The human seminoma‐derived TC am‐2 cell line was exposed to activin A (24 h), and target transcripts were measured by qRT ‐ PCR ( n = 4). ELISA ( n = 4) and gelatin zymography ( n = 3) showed changes in protein level and enzyme activity, respectively. Results (i) Cytoplasmic CXCL 12 was detected in Sertoli and other somatic cells, including those surrounding seminoma cells. Anti‐ CCL 17 labelled only the cytoplasm of Sertoli cells surrounding GCNIS , while anti‐ MMP 2 and anti‐ MMP 9 labelled germline and epithelial‐like cells in normal and neoplastic testes. (ii) Exposing TC am‐2 cells to activin A (50 ng/ mL ) elevated MMP 2 and MMP 9 transcripts (fourfold and 30‐fold), while only MMP 2 protein levels were significantly higher after activin A (5 ng/ mL and 50 ng/ mL ) exposure. Importantly, gelatin zymography revealed activin A increased production of activated MMP2. Discussion Detection of CCL 17 only in GCNIS tumours may reflect a change in Sertoli cell phenotype to a less mature state. Stimulation of MMP 2 activity by activin A in TC am‐2 cells suggests activin influences TGCT by modulating the tumour niche. Conclusion This knowledge provides a basis for understanding how physiological changes that influence activin/ TGF ‐β superfamily signalling may alter germ cell fate.