Abstract 2254: Pro-angiogenic CXCL8 signalling underpins microenvironment-induced relapse to anti-androgen therapy of prostate cancer

Abstract

Abstract Introduction and Objectives: Bicalutamide induces a rapid and sustained hypoxia in growing LNCaP prostatic tumors. This treatment-induced hypoxia (>14 days) increases synthesis of the CXC-chemokine Interleukin-8(CXCL8) co-incident with increasing oxygen tension and the restoration of tumor vascularisation1. Consequently, our current hypothesis is that hypoxia induced CXCL8 signalling underpins resistance to bicalutamide-treated tumors and facilitates the adoption of more aggressive cancer cells. Results: Culturing of LNCaP (PTEN null) and 22Rv1 (PTEN wt) cells under hypoxic conditions (mimicking a bicalutamide-treated tumor microenvironment) increased transcriptional activity of the androgen receptor (AR), Hypoxia-Inducible Factor (HIF-1) and NF-kB, inducing expression of downstream target genes associated with survival, metabolic adaptation and angiogenesis. The magnitude and duration of these responses was greater in PTEN-null LNCaP cells. Experiments were conducted to determine how bicalutamide-mediated inhibition of AR signalling would affect hypoxia-induced potentiation of AR, HIF and NF-kB transcription. Bicalutamide or an AR-targeted siRNA failed to reduce expression of pro-angiogenic, pro-survival and metabolism-associated genes. However, concurrent knockdown of both HIF-1 and NF-kB activity using gene-targeted siRNA's repressed expression of these hypoxia-induced disease-progressing genes. Amongst the genes up-regulated in hypoxic and PTEN-deficient LNCaP cells was expression of CXCL8 and its receptors, CXCR1 and CXCR2, which we have previously shown to be regulated by HIF-1 and NF-kB activity. Potentiation of CXCL8 signalling under hypoxia was found to sustain AR, HIF and NF-kB transcription. Combination of bicalutamide with the proteasome inhibitor bortezomib attenuated hypoxia-induced AR expression and transcriptional activity and decreased HIF-1 activity but failed to reduce NF-kB activity. In contrast, only direct inhibition of CXCL8 signalling was shown to repress activity of AR, HIF-1 and NF-kB in hypoxic LNCaP cells and increased LNCaP cells sensitivity to bicalutamide. Conclusion: Our data suggests that targeting CXCL8 signalling may therefore be a relevant strategy to enhance tumor response to anti-androgen therapies. Inhibiting signalling of this chemokine attenuates key transcriptional approaches induced in the hypoxic microenvironment of bicalutamide-treated tumors. Moreover, inhibition of CXCL8 signalling may prevent the re-vascularization previously observed in hypoxic LNCaP tumors. Citation Format: Melanie McKechnie, Pamela J. Maxwell, David J. J. Waugh. Pro-angiogenic CXCL8 signalling underpins microenvironment-induced relapse to anti-androgen therapy of prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2254. doi:10.1158/1538-7445.AM2014-2254