IL-1ß induces p62/SQSTM1 and represses androgen receptor expression in prostate cancer cells.

Chronic inflammation is associated with advanced prostate cancer (PCa), although the mechanisms governing inflammation-mediated PCa progression are not fully understood. PCa progresses to an androgen independent phenotype that is incurable. We previously showed that androgen independent, androgen receptor negative (AR(-) ) PCa cell lines have high p62/SQSTM1 levels required for cell survival. We also showed that factors in the HS-5 bone marrow stromal cell (BMSC) conditioned medium can upregulate p62 in AR(+) PCa cell lines, leading us to investigate AR expression under those growth conditions. In this paper, mRNA, protein, and subcellular analyses reveal that HS-5 BMSC conditioned medium represses AR mRNA, protein, and nuclear accumulation in the C4-2 PCa cell line. Using published gene expression data, we identify the inflammatory cytokine, IL-1ß, as a candidate BMSC paracrine factor to regulate AR expression and find that IL-1ß is sufficient to both repress AR and upregulate p62 in multiple PCa cell lines. Immunostaining demonstrates that, while the C4-2 population shows a primarily homogeneous response to factors in HS-5 BMSC conditioned medium, IL-1ß elicits a strikingly heterogeneous response; suggesting that there are other regulatory factors in the conditioned medium. Finally, while we observe concomitant AR loss and p62 upregulation in IL-1ß-treated C4-2 cells, silencing of AR or p62 suggests that IL-1ß regulates their protein accumulation through independent pathways. Taken together, these in vitro results suggest that IL-1ß can drive PCa progression in an inflammatory microenvironment through AR repression and p62 induction to promote the development and survival of androgen independent PCa.

TISSUE ENGINEERING PERFUSABLE CANCER MODELS.

The effect of fluid flow on cancer progression is currently not well understood, highlighting the need for perfused tumor models to close this gap in knowledge. Enabling biological processes at the cellular level to be modeled with high spatiotemporal control, microfluidic tumor models have demonstrated applicability as platforms to study cell-cell interactions, effect of interstitial flow on tumor migration and the role of vascular barrier function. To account for the multi-scale nature of cancer growth and invasion, macroscale models are also necessary. The consideration of fluid dynamics within tumor models at both the micro- and macroscopic levels may greatly improve our ability to more fully mimic the tumor microenvironment.

Monomer-dimer equilibrium of the pSC101 RepA protein.

The pSC101 RepA protein, which is required for plasmid DNA replication, but is inhibitory to replication at high concentration, has been found in both monomeric and dimeric forms. While RepA monomers bind to direct repeat iterons near the pSC101 replication origin, dimers bind to sequences that autoregulate RepA synthesis. We investigated the solution properties of purified RepA protein by analytical ultracentrifugation analysis, and found that RepA exists in Escherichia coli cells in a monomer-dimer equilibrium (Kd = 4 microM), and, moreover, that RepA is primarily in the monomeric form at the concentration (500 molecules per cell; 2 microM) we found by Western blot analysis to occur in cells carrying replicating wild-type pSC101 plasmids. However, at concentrations inhibitory to pSC101 DNA replication, the majority of RepA molecules exist as dimers. Our findings provide experimental support for the proposal that the equilibrium between monomer and dimer forms of RepA has a key role in determining its effect on the replication of pSC101.