Interferon γ inhibits growth of human pancreatic carcinoma cells via caspase-1 dependent induction of apoptosis
Interferon γ inhibits growth of human pancreatic carcinoma cells via caspase-1 dependent induction of apoptosis
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DOI：
10.1136/gut.49.2.251
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发表时间：
2001-08-01
期刊：
影响因子：
24.5
通讯作者：
Rosewicz, S
中科院分区：
文献类型：
--
作者：
Detjen, KM;Farwig, K;Rosewicz, S
背景和目的：胰腺癌预后不良的部分原因是对广谱凋亡刺激的抵抗。为了确定完整的促凋亡途径的潜在临床意义，我们的特点是干扰素γ（IFN-γ）对人胰腺癌细胞的生长和生存的影响。方法-IFN-γ受体的表达和信号转导进行了研究，通过逆转录-聚合酶链反应（RT-PCR），免疫沉淀，蛋白质印迹分析，和反式激活试验。对细胞生长和存活的影响进行了评估，在细胞数量，集落形成，细胞周期分析，DNA片段化，和聚（ADP核糖）聚合酶（PARP）cleavage.Results -所有四个胰腺癌细胞系检查表达功能IFN-γ受体和下游效应，包括假定的肿瘤抑制因子干扰素调节因子1（IRF-1）。IFN-γ处理显著抑制胰腺癌细胞的锚定依赖性和非锚定依赖性生长。细胞周期分析显示亚二倍体细胞提示凋亡，这是证实了DNA片段化和PARP裂解。细胞凋亡诱导和生长抑制的时间和剂量依赖性密切相关，确定细胞凋亡是主要的，如果不是唯一的，负责生长抑制的机制。细胞凋亡之前的procaspase-1的上调，并伴随着蛋白水解激活。此外，半胱天冬酶抑制剂z-vad-festival完全防止IFN-γ介导的apoptosis.Conclusions -这些结果确定了一个完整的胰腺癌细胞的促凋亡途径，并建议IRF-1和/或procaspase-1可能代表潜在的治疗目标，以进一步探讨。
Background and aims - The poor prognosis of pancreatic cancer is partly due to resistance to a broad spectrum of apoptotic stimuli. To identify intact proapoptotic pathways of potential clinical relevance, we characterised the effects of interferon gamma (IFN-gamma) on growth and survival in human pancreatic cancer cells.Methods - IFN-gamma receptor expression and signal transduction were examined by reverse transcriptase-polymerase chain reaction (RT-PCR), immunoprecipitation, western blot analysis, and transactivation assays. Effects on cell growth and survival were evaluated in terms of cell numbers, colony formation, cell cycle analysis, DNA fragmentation, and poly(ADP ribose) polymerase (PARP) cleavage.Results - All four pancreatic cancer cell lines examined expressed functional IFN-gamma receptors and downstream effecters, including the putative tumour suppressor interferon regulatory factor 1 (IRF-1). IFN-gamma treatment profoundly inhibited anchorage dependent and independent growth of pancreatic cancer cells. Cell cycle analyses revealed subdiploid cells suggesting apoptosis, which was confirmed by demonstration of DNA fragmentation and PARP cleavage. Time and dose dependency of apoptosis induction and growth inhibition correlated closely, identifying apoptosis as the main, if not exclusive, mechanism responsible for growth inhibition. Apoptosis was preceded by upregulation of procaspase-1 and accompanied by proteolytic activation. Furthermore, the caspase inhibitor z-vad-fmk completely prevented IFN-gamma mediated apoptosis.Conclusions - These results identify an intact proapoptotic pathway in pancreatic cancer cells and suggest that IRF-1 and/or procaspase-1 may represent potential therapeutic targets to be further explored.