STAT1 modification improves therapeutic effects of interferons on lung cancer cells.
STAT1 modification improves therapeutic effects of interferons on lung cancer cells.
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STAT1修饰提高干扰素对肺癌细胞的治疗效果
DOI：
10.1186/s12967-015-0656-0
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发表时间：
2015-09-08
影响因子：
7.4
通讯作者：
Chen C
中科院分区：
文献类型：
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作者：
Chen J;Zhao J;Chen L;Dong N;Ying Z;Cai Z;Ji D;Zhang Y;Dong L;Li Y;Jiang L;Holtzman MJ;Chen C
背景干扰素（IFN）具有抗肿瘤增殖、促凋亡和免疫调节作用。然而，IFN的临床效用是有限的毒性和药代动力学，使其难以达到持续的治疗水平，特别是在实体tumorsMethodsSignal Transducer and Activator of Transcription 1（STAT 1）或一个修改后的STAT 1（指定STAT 1-CC），这是高反应的IFN过表达肺癌SPC-A-1和H1299细胞使用慢病毒载体。使用增强的绿色荧光蛋白（EGFP）表达监测转导效率。转导后，用干扰素-γ（IFN-γ）或干扰素-β（IFN-β）处理细胞，并使用细胞计数试剂盒-8和transwell小室测定法监测细胞增殖、迁移和侵袭力，并通过流式细胞术使用膜联蛋白V检测监测细胞凋亡。此外，使用蛋白质印迹法测定STAT 1、STAT 1 Tyr-701磷酸化（pSTAT 1）、纤连蛋白和β-连环蛋白的水平。在IFN-γ刺激的情况下，S100 A4，增殖细胞核抗原（PCNA），和c-fos的表达水平也进行了测定。结果我们发现，STAT 1或STAT 1-CC的表达增强IFN-γ和IFN-β对人肺癌细胞增殖，迁移和侵袭的抑制作用。此外，STAT 1和STAT 1-CC的表达引起pSTAT 1的增加和纤连蛋白和β-连环蛋白水平的降低。STAT 1-CC对IFN-γ诱导的pSTAT 1表达的抑制作用较STAT 1强，对S100 A4、PCNA、c-fos表达的抑制作用较STAT 1强。本研究结果提示，IFN和STAT 1-CC联合治疗可能是未来肺癌临床治疗的一种可行方法。
BackgroundInterferons (IFNs) have potent anti-proliferative, pro-apoptotic, and immunomodulatory activities against cancer. However, the clinical utility of IFNs is limited by toxicity and pharmacokinetics making it difficult to achieve sustained therapeutic levels especially in solid tumors.MethodsSignal Transducer and Activator of Transcription 1 (STAT1) or a modified STAT1 (designated STAT1-CC) that is hyper-responsive to IFN were overexpressed in lung cancer SPC-A-1 and H1299 cells using lentiviral vectors. Transduction efficiency was monitored using enhanced green fluorescent protein (EGFP) expression. After transduction, cells were treated with interferon-gamma (IFN-γ) or interferon-beta (IFN-β) and monitored for cell proliferation, migration, and invasiveness using Cell Counting Kit-8 and transwell chamber assays and for apoptosis using Annexin V detection by flow cytometry. In addition, levels of STAT1, STAT1 Tyr-701 phosphorylation (pSTAT1), fibronectin, and β-catenin were determined using western blotting. In the case of IFN-γ stimulation, levels of S100A4, proliferating cell nuclear antigen (PCNA), and c-fos expression were also determined.ResultsWe found that expression of STAT1 or STAT1-CC enhanced the effect of IFN-γ and, IFN-β on inhibition of human lung cancer cell proliferation, migration and invasiveness. Moreover, STAT1 and STAT1-CC expression caused increases in pSTAT1 and decreases in fibronectin and β-catenin levels. STAT1-CC showed increased effects compared to STAT1 on IFN-γ induced pSTAT1 and down-regulation of S100A4, PCNA, and c-fos levels.ConclusionThe results show that STAT1-CC exhibited more strength in improving the antitumor response of IFNs in lung cancer cells. Results from this study suggest that combined treatment of IFNs and STAT1-CC might be a feasible approach for the clinical management of lung cancer in the future.