Armstrong Chris W D, Maxwell Pamela J, Ong Chee Wee, Redmond Kelly M, McCann Christopher, Neisen Jessica, Ward George A, Chessari Gianni, Johnson Christopher, Crawford Nyree T, LaBonte Melissa J, Prise Kevin M, Robson Tracy, Salto-Tellez Manuel, Longley Daniel B, Waugh David J J
Movember Centre of Excellence, Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast, Northern Ireland.
Astex Pharmaceuticals, Cambridge, UK.
Oncotarget. 2016 Feb 16;7(7):7885-98. doi: 10.18632/oncotarget.6955.
PTEN loss is prognostic for patient relapse post-radiotherapy in prostate cancer (CaP). Infiltration of tumor-associated macrophages (TAMs) is associated with reduced disease-free survival following radical prostatectomy. However, the association between PTEN loss, TAM infiltration and radiotherapy response of CaP cells remains to be evaluated. Immunohistochemical and molecular analysis of surgically-resected Gleason 7 tumors confirmed that PTEN loss correlated with increased CXCL8 expression and macrophage infiltration. However PTEN status had no discernable correlation with expression of other inflammatory markers by CaP cells, including TNF-α. In vitro, exposure to conditioned media harvested from irradiated PTEN null CaP cells induced chemotaxis of macrophage-like THP-1 cells, a response partially attenuated by CXCL8 inhibition. Co-culture with THP-1 cells resulted in a modest reduction in the radio-sensitivity of DU145 cells. Cytokine profiling revealed constitutive secretion of TNF-α from CaP cells irrespective of PTEN status and IR-induced TNF-α secretion from THP-1 cells. THP-1-derived TNF-α increased NFκB pro-survival activity and elevated expression of anti-apoptotic proteins including cellular inhibitor of apoptosis protein-1 (cIAP-1) in CaP cells, which could be attenuated by pre-treatment with a TNF-α neutralizing antibody. Treatment with a novel IAP antagonist, AT-IAP, decreased basal and TNF-α-induced cIAP-1 expression in CaP cells, switched TNF-α signaling from pro-survival to pro-apoptotic and increased radiation sensitivity of CaP cells in co-culture with THP-1 cells. We conclude that targeting cIAP-1 can overcome apoptosis resistance of CaP cells and is an ideal approach to exploit high TNF-α signals within the TAM-rich microenvironment of PTEN-deficient CaP cells to enhance response to radiotherapy.
PTEN缺失对前列腺癌(CaP)患者放疗后复发具有预后价值。肿瘤相关巨噬细胞(TAM)浸润与根治性前列腺切除术后无病生存期缩短相关。然而,PTEN缺失、TAM浸润与CaP细胞放疗反应之间的关联仍有待评估。对手术切除的Gleason 7级肿瘤进行免疫组化和分子分析证实,PTEN缺失与CXCL8表达增加及巨噬细胞浸润相关。然而,PTEN状态与CaP细胞其他炎症标志物的表达,包括TNF-α,没有明显相关性。在体外,将巨噬细胞样THP-1细胞暴露于从照射后的PTEN缺失CaP细胞收集的条件培养基中可诱导其趋化,CXCL8抑制可部分减弱该反应。与THP-1细胞共培养导致DU145细胞放射敏感性适度降低。细胞因子分析显示,无论PTEN状态如何,CaP细胞均组成性分泌TNF-α,且照射可诱导THP-1细胞分泌TNF-α。THP-1来源的TNF-α增加了CaP细胞中NFκB的促生存活性,并提高了包括细胞凋亡抑制蛋白-1(cIAP-1)在内的抗凋亡蛋白的表达,用TNF-α中和抗体预处理可减弱这种作用。用新型IAP拮抗剂AT-IAP处理可降低CaP细胞中基础和TNF-α诱导的cIAP-1表达,将TNF-α信号从促生存转变为促凋亡,并增加与THP-1细胞共培养的CaP细胞的放射敏感性。我们得出结论,靶向cIAP-1可克服CaP细胞的凋亡抗性,是利用PTEN缺陷型CaP细胞富含TAM的微环境中高TNF-α信号来增强放疗反应的理想方法。