siRNA沉默SOCS1表达增强IFN-α2a-NGR的抗肿瘤效应

SOCS1 knockdown sensitize anti-tumor activity of IFN-α2a-NGR

目的:研究肿瘤血管导向性干扰素IFN-α2a-NGR抗肿瘤效应的分子机制,发现影响干扰素敏感性的关键信号分子,提高耐药肿瘤细胞敏感性。方法:培养IFNAR高表达细胞株A549、IFNAR低表达细胞株MKN-45,通过MTT检测IFN-α2a-NGR的抗细胞增殖活性,流式细胞术(FCM)、Western blot检测IFN-α2a-NGR处理前后STAT1、p-STAT1、p53、OAS与SOCS1的表达变化,合成siRNA靶向干涉SOCS1。结果:IFN-α2a-NGR刺激后,A549中,p-STAT1、p53、OAS与SOCS1表达上调;MKN-45中P53、OAS无显著变化,SOCS1显著上调。靶向干涉SOCS1后,MKN-45对IFN-α2a-NGR的敏感性增加[抑制率从(14.69±1.05)%提高到(36.97±2.05)%]。结论:IFN-α2a-NGR与IFN-α2a在细胞和分子水平的效应基本一致,p-STAT1、p53与SOCS1可影响细胞对干扰素的敏感性,通过siRNA沉默技术可提高耐药细胞株敏感性,为IFN-α2a-NGR的进一步研发奠定了基础。

AIM：Search for key molecules to influence the tumor-targeted IFN-α2a-NGR anti-tumor sensitivity through signaling pathway study.Try to enhance the antitumor efficacy of IFN-α2a-NGR.METHODS：MTT method was used to determine the growth inhibitory effects of IFN-α2a-NGR on A549 and MKN-45 cells;Flow cytometry and Western blot were employed to detect the expression of STAT1,p-STAT1,p53,OAS and SOCS1;SOCS1 gene knock down was carried out by synthesized siRNA.RESULTS：When stimulated with IFN-α2a-NGR,the increased expression of STAT1,p-STAT1,p53,OAS and SOCS1 were observed in A549 cells,but only SOCS1 was notably increased in MKN-45 cells.The proliferation inhibition ability of MKN-45 to IFN-α2a-NGR was promoted by SOCS1 knocking down.（the inhibition rate was enhanced from 14.69%±1.05% to 36.97%±2.05%）.CONCLUSION：This study has further demonstrated that there were no differences on antitumor effects between IFN-α2a-NGR and IFN-α2a on cell or molecular level.Besides interferon-α receptor （IFNAR） which has been demonstrated before,p-STAT1,p53 and SOCS1 were important determinants of tumor resistance to IFNs therapy.The antitumor efficacy of IFN-α2a-NGR can be enhanced by RNA interference.These results might be helpful for the further development of IFN-α2a-NGR.