PTC-209靶向抑制Bmi1对头颈鳞癌细胞的体外抗癌作用研究

【】目的：检测Bmi1(B lymphoma Mo-MLV insertion region 1)小分子化学抑制剂PTC-209对头颈鳞癌细胞中Bmi1的表达抑制作用,并探讨其对体外头颈鳞癌细胞生物表型的影响。方法：对头颈鳞癌细胞系Cal27和FaDu予以PTC-209处理,通过蛋白质免疫印迹、实时定量逆转录聚合酶链反应检测PTC-209干预后细胞内Bmip1的表达变化；通过MTT、Transwell、划痕试验等分析PTC-209处理对头颈鳞癌细胞增殖、迁移和侵袭的影响；应用克隆形成、肿瘤球培养、流式分选等实验分析PTC-209对头颈鳞癌细胞系中肿瘤干细胞亚群的影响。结果：PTC-209能显著下调头颈鳞癌细胞中Bmi1的表达,并具有浓度和时间依赖效应(P＜0.05); PTC-209体外干预可明显抑制细胞增殖、侵袭和迁移能力,增强细胞对顺铂和5-FU的药物敏感性；PTC-209能显著降低细胞克隆形成率、肿瘤球形成以及ALDH₊亚群细胞比例。结论：PTC-209能在体外抑制头颈鳞癌细胞中Bmi1的表达,具有抑制细胞增殖,侵袭迁移和降低肿瘤干细胞亚群比例等抗肿瘤效应。

Therapeutic targeting of Bmi1 with PTC-209 in head neck squamous cell carcinoma in vitro

Objective: In the present study, we aimed to investigate the small-molecule Bmi1 inhibitor, PTC-209, for its effects on endogenous Bmi1 expression and cell behaviors in head neck squamous cell carcinoma (HNSCC) cell lines in vitro. Methods: The HNSCC cell lines Cal27 and FaDu were treated with PTC-209 in vitro and the expression change of Bmi1 was detected by western blot and real-time RT-PCR assays. The changes of cell proliferation, invasion and migration were analyzed by MTT, transwell and wound-healing assays following PTC-209 treatment. The cancer stem cell (CSC) subpopulations in Cal27 and FaDu cells were further assayed by tumorsphere formation, colony-forming and flow cytometry assays. Results: Bmi1 mRNA and protein levels were significantly decreased in Cal27 and FaDu cells treated with PTC-209 in both dose- and time dependent manners. PTC-209 inhibited cell proliferation, migration and invasion, and had synergic anti-cancer effects when combined with cisplatin and 5-FU. Moreover, colony formation, sphere formation, and ALDH₊ subpopulation were significantly reduced upon PTC-209 exposure. Conclusion: PTC-209 treatment induces Bmi1 reduction in HNSCC cell lines in vitro, and shows potent anti-tumor activity with reduced cell proliferation, migration, invasion and CSC subpopulation.