Projects per year
Abstract
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. CXCL12, CCL17, MMP2 and MMP9 were investigated. (2) The human seminoma-derived TCam-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 CXCL12 was detected in Sertoli and other somatic cells, including those surrounding seminoma cells. Anti-CCL17 labelled only the cytoplasm of Sertoli cells surrounding GCNIS, while anti-MMP2 and anti-MMP9 labelled germline and epithelial-like cells in normal and neoplastic testes. (ii) Exposing TCam-2 cells to activin A (50 ng/mL) elevated MMP2 and MMP9 transcripts (fourfold and 30-fold), while only MMP2 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 CCL17 only in GCNIS tumours may reflect a change in Sertoli cell phenotype to a less mature state. Stimulation of MMP2 activity by activin A in TCam-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.
Original language | English |
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Pages (from-to) | 31-41 |
Number of pages | 11 |
Journal | Andrology |
Volume | 7 |
Issue number | 1 |
DOIs | |
Publication status | Published - 1 Jan 2019 |
Keywords
- activin A
- chemokine
- matrix metalloproteinase
- neoplasia
- testicular germ cell tumours
Projects
- 3 Finished
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Activin Control of the Male Germline for Reproductive Health
National Health and Medical Research Council (NHMRC) (Australia)
1/01/15 → 31/12/18
Project: Research
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Activin and its binding protein, follistatin regulate functions of the epididymis and vas deferens
Hedger, M., Loveland, K., Meinhardt, A. & de Kretser, D.
National Health and Medical Research Council (NHMRC) (Australia)
1/01/14 → 31/12/16
Project: Research
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NHMRC Research Fellowship
National Health and Medical Research Council (NHMRC) (Australia)
1/01/03 → 31/12/19
Project: Research