Ad-FGF-Trap对头颈部鳞癌细胞的抑制及其与西妥昔单抗的联合作用

Inhibition Effect of Ad-FGF-Trap on Head and Neck Squamous Cell Carcinoma Cells and Its Combination Therapy with Cetuximab

西妥昔单抗的耐药问题在头颈部鳞癌(head and neck squamous cell carcinoma,HNSCC)的治疗过程中较为突出,成纤维生长因子受体1(fibroblast growth factor receptor 1,FGFR1)旁路活化是一个重要原因。在抑制FGFR1活性方面,已有的FGFR小分子抑制剂副作用较大,"诱饵受体"蛋白可能具有优势。该工作通过分子克隆方法构建了腺病毒穿梭质粒pDC316-FGFR1 Ig2+Ig3用以表达人FGF的"诱饵受体",利用AdMax;腺病毒系统包装、扩增、纯化获得了高滴度的Ad-FGF-Trap重组腺病毒。Ad-FGF-Trap感染HNSCC细胞CAL 27、WSU-HN6后,可显著抑制细胞增殖、迁移、侵袭能力。CAL 27细胞感染Ad-FGF-Trap后发生凋亡,同时PI3K/MAPK信号通路受到抑制,PARP剪切增加。WSU-HN6细胞感染Ad-FGF-Trap后,p27、p53、PCNA等周期相关蛋白表达量增加,细胞发生G;/S期阻滞,但EGFR通路代偿性激活,显示出FGFR1和EGFR两个信号通路之间有重要的相关性。利用CAL 27细胞进行的裸鼠体内实验表明Ad-FGF-Trap可抑制肿瘤生长。进一步研究表明,Ad-FGF-Trap联合西妥昔单抗,可更有效地抑制FGFR1及EGFR同时高表达的WSU-HN6细胞增殖,显示出联合用药将获得更佳疗效。该研究表明可表达"诱饵受体"的Ad-FGF-Trap能够为HNSCC的靶向治疗提供新的思路。

The bypass activation of FGFR1(fibroblast growth factor receptor 1)is one of the major reasons for the common resistance to Cetuximab in the treatment of HNSCC(head and neck squamous cell carcinoma).“Decoy receptor”may have advantages over small molecule inhibitor in FGFR1 inactivation with regard to side effects.In this work,a recombinant shuttle plasmid pDC316-FGFR1 Ig2+Ig3 was constructed by genetic engineering for the expression of“Decoy receptor”.High-titer recombinant adenovirus Ad-FGF-Trap was obtained after packaging,amplification and purification using AdMax;adenovirus system.The proliferation,migration,invasion of human HNSCC cells CAL 27 and WSU-HN6 were significantly inhibited by infection of Ad-FGF-Trap.Apoptosis of CAL 27 cells occurred when treated with Ad-FGF-Trap,accompanied by down-regulation of PI3K/MAPK signaling pathway and up-regulation of cleaved PARP.G;/S-arrest was observed for WSU-HN6 cells treated with Ad-FGF-Trap,accompanied with up-regulation of cyclin-dependent proteins p27,p53 and PCNA,whereas enhanced compensatory activation of EGFR pathway.Ad-FGF-Trap inhibited tumor growth of CAL 27 xenografts in nude mice in vivo.Furthermore,Ad-FGF-Trap combined with Cetuximab more effectively inhibited the proliferation of WSU-HN6 cells which had high FGFR1 and EGFR expression.The current study of“decoy receptor”-expressing Ad-FGF-Trap provides a new strategy for the targeted therapy of HNSCC.