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Effect of Tumor Conditioned Media from Prostate Cancer Cells on Fibroblast Cell Morphology, Viability and Gene Expression
Author(s) -
Kaur Sukhneeraj Pal,
Cummings Brian S.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.566.15
Subject(s) - du145 , stromal cell , cancer cell , tumor microenvironment , cancer research , prostate cancer , viability assay , cancer , biology , extracellular matrix , cell growth , metastasis , carcinogenesis , microbiology and biotechnology , cell , chemistry , lncap , biochemistry , genetics , tumor cells
Cancer is a heterogeneous collection of neoplastic cells, surrounded by non‐malignant, stromal cells, and extracellular matrix (ECM), among other constituents. Cross‐talk between tumor and stromal cells forms a dynamic tumor microenvironment (TME) that mediates cell growth, proliferation, and metastasis. The TME can act to inhibit cancer growth and proliferation during the initial stages of carcinogenesis. However, cancer cells are able to evade these inhibitory signals, and subsequently reshape the TME, making it more amenable to their own growth. This gives rise to numerous cancer‐associated stromal cells, including cancer‐associated fibroblasts (CAFs). CAFs further modulate the TME by regulating the release of growth signals, cell death, ECM remodeling, angiogenesis, and epithelial‐mesenchymal transition. To study the ability of prostate cancer to alter the TME in vitro , we exposed human bone fibroblasts (HS‐5 cells) to tumor conditioned media isolated from a human neuroendocrine prostate cancer cell line (DU145) and assessed changes in cell morphology, viability (using MTT assay) and the expression of Group IIA secretory phospholipase A 2 (PLA2G2A), as well as its regulatory proteins, glypican‐1 (GPC‐1) and Group IVA phospholipase A 2 (PLA2G4A), using qPCR. Exposure of HS‐5 cells to TCM from DU145 cells altered cellular morphology to a more mesenchymal phenotype typically seen in active myofibroblasts. TCM also decreased MTT staining in a time‐dependent manner that was independent of the presence of serum in the TCM. Changes in cell morphology and viability correlated to increases in the expression of PLA2G2A, GPC‐1 and PLA2G4A. The effect of TCM was time‐ and serum‐dependent, and was differential for each gene studied. Alteration in cellular morphology and increases in PLA2G2A, GPC‐1 and PLA2G4A in HS‐5 cells correlated to increases in the expression of fibroblast activation protein‐α, which is a CAF marker. To elucidate the mechanisms mediating changes in HS‐5 cells exposed to TCM, we investigated the expression of GLi1, a transcription factor involved in the activation of the sonic hedgehog signaling pathway. We found that TCM increased GLi1 mRNA levels in HS‐5 cells, with greater increases being observed using TCM containing serum. Collectively, these data demonstrate that media from DU145 cells can induce morphological and genotypic changes in human fibroblasts similar to those observed in CAFs. As such, this model holds promise for identifying molecular mechanisms by which prostate cancer cells alter the TME and induce prostate cancer cell growth. Support or Funding Information National Institute of Biomedical Imaging and Bioengineering NIBIB (EB0160100)Department of Defense Prostate Cancer Research Program Idea Development Award (PC150431 GRANT11996600) This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .