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目的:探讨受体酪氨酸激酶anexelekto(Axl)在鼻咽癌(nasopharyngeal carcinoma,NPC)中的表达及意义。方法:采用免疫组化法检测78例NPC和32例鼻咽黏膜慢性炎中Axl的表达,分析Axl蛋白表达与NPC患者临床参数的相关性。常规培养NPC细胞,免疫荧光法检测不同分化NPC细胞系CNE1、CNE2Z及C666-1中Axl的蛋白表达情况。应用Axl特异性抑制剂TP-0903处理CNE1和C666-1细胞,CCK-8实验检测细胞的活力,流式细胞术检测细胞周期的分布,q PCR检测Axl和增殖细胞核抗原(PCNA)的mRNA表达,Western blot检测Axl及p-Axl蛋白的表达。结果:Axl蛋白定位于胞膜和胞质。NPC中Axl高表达阳性率显著高于鼻咽黏膜慢性炎(P<0.01)。Axl高表达与患者年龄、性别及M分期无关,与临床分期、T分期和N分期呈正相关(P<0.05)。Axl在高分化CNE1细胞中低表达,在低分化CNE2Z细胞和未分化C666-1细胞中表达水平明显增高。TP-0903呈浓度和时间依赖性抑制NPC细胞的活性,2 nmol/L TP-0903即具有显著抑制效应,能阻滞细胞周期于G0期,在降低Axl活性的同时也显著抑制PCNA的表达。结论:Axl高表达可促进NPC的临床进展;TP-0903显著抑制NPC细胞的增殖,提示Axl可能在NPC靶向治疗中具有一定的价值。
Objective: To investigate the expression and significance of anexelekto (Axl), a receptor tyrosine kinase in nasopharyngeal carcinoma (NPC). Methods: The expression of Axl in 78 cases of NPC and 32 cases of chronic nasopharyngeal mucositis was detected by immunohistochemistry, and the correlation between Axl protein expression and clinical parameters of NPC was analyzed. The NPC cells were routinely cultured, and Axl protein expression in different differentiated NPC cell lines CNE1, CNE2Z and C666-1 was detected by immunofluorescence. CNE1 and C666-1 cells were treated with Ax-specific inhibitor TP-0903. The viability of cells was detected by CCK-8 assay. The distribution of cell cycle was detected by flow cytometry. The mRNA expression of Axl and proliferating cell nuclear antigen (PCNA) Western blot was used to detect Axl and p-Axl protein expression. Results: Axl protein localized in the plasma membrane and cytoplasm. The positive rate of Axl overexpression in NPC was significantly higher than that in nasopharyngeal mucosa (P <0.01). Axl expression was not related with age, sex and M stage, but positively correlated with clinical stage, T stage and N stage (P <0.05). Axl was overexpressed in well-differentiated CNE1 cells and was significantly up-regulated in poorly differentiated CNE2Z cells and undifferentiated C666-1 cells. TP-0903 inhibited the activity of NPC cells in a concentration- and time-dependent manner. 2-nmol / L TP-0903 had a significant inhibitory effect and blocked the cell cycle at G0 phase. It decreased Axl activity and significantly inhibited PCNA expression. Conclusion: Axl overexpression can promote the clinical progress of NPC. TP-0903 significantly inhibits the proliferation of NPC cells, suggesting that Axl may be of value in NPC targeted therapy.