姜黄素衍生物FM0807体外抗鼻咽癌作用及其机制的研究

The Anti-nasopharyngeal Carcinoma Activity and Mechanism of Curcumin Derivative FM0807

目的观察姜黄素衍生物FM0807的抗鼻咽癌细胞增殖效应,并探讨其机制。方法 MTT法、台盼蓝排染计数法、集落形成试验检测FM0807对人鼻咽癌细胞CNE1及CNE2增殖的抑制作用。吉姆萨染色、Hoechest33342/PI双染检测FM0807对鼻咽癌细胞凋亡及形态的影响。免疫共沉淀研究药物对Hsp90的分子伴侣功能的影响。Western blot检测FM0807处理后鼻咽癌细胞相关分子蛋白含量变化。结果 (1)FM0807呈剂量依赖性地抑制鼻咽癌细胞增殖、诱导其凋亡并引起细胞形态改变;(2)FM0807能够抑制AKT和ERK1/2与Hsp90的结合,影响其信号转导;(3)FM0807引起鼻咽癌细胞P-AKT、P-ERK1/2、NF-κB、Cdk4、MMP9蛋白含量下降,而Bax增多,还能对抗表皮生长因子介导的P-ERK1/2信号的激活。结论 FM0807抗鼻咽癌细胞作用的机制可能是通过抑制Hsp90与AKT和ERK1/2的结合,从而抑制PI3K/AKT与MAPK/ERK信号传导通路。

Objective To investigate the antiproliferative effect of curcumin derivative FM0807 on Nasopharyngeal Carcinoma（NPC） cells and its mechanism. Methods NPC cell lines CNE1 and CNE2 were treated with FM0807, and MTT, Trypan Blue staining and colony formation assays were used to e- valuate the effect of the drugs. Giemsa and Hoechest33342/PI were applied to determine apoptosis and shape of NPC cells with FM0807 treatment. Co-immunoprecipitation assays were used to analysis the chaperone function of Hsp90. We applied Western blot to show relative proteins expression after FM0807 incubation. At the same time, we analysis whether FM0807 affect MAPK/ERK survival signa- ling pathway mediated by EGF. Results Assays showed inhibition of proliferation in NPC cells with FM0807 incubation in a dose-dependent way. FM0807 treatment caused apoptosis and changed shape of NPC cells. Immunoprecipitation assays showed FM0807 affected down-stream signal via disturbed Hsp90 bind to its client（AKT and ERK1/2）. Western blot analyses showed decreased P-AKT,P-ERK1/2,NF- ΚB,Cdk4,MMP9 and increased Bax in NPC cells. FM0807 also can downregulation of P-ERK1/2 media- ted by EGF. Conclusion FM0807 can inhibit the proliferation of NPC cells, and the mechanism may be FM0807 downregulation of PI3K/AKT and MAPK/ERK signaling pathways via inhibition of Hsp90 bind to AKT and ERK1/2.