摘要
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。
AIM:To explore the expression and significance of receptor tyrosine kinase anexelekto(Axl)in nasopharyngeal carcinoma(NPC).METHODS:Immunohistochemistry was used to detect the Axl protein expression of78patients with NPC and32patients with nasopharyngeal chronic inflammation(NPI).The correlations between the Axl pro-tein levels and the clinical parameters of NPC patients were analyzed.NPC cells were cultured in vitro,and the expression of Axl in well differentiated CNE1cells,poorly-differentiated CNE2Z cells and undifferentiated C666-1cells was detected by immunofluorescence staining.After treatment of the CNEland C666-1cells with Axl specific inhibitor TP-0903,CCK-8assay was used to detect cell viability,flow cytometry was adopted to analyze the cell cycle distribution,qPCR was used to examine the mRNA levels of Axl and proliferating cell nuclear antigen(PCNA),and Western blot was used to examine the protein expression of Axl and p-Axl.RESULTS:Axl protein was localized in the cell membrane and cytoplasm.The rate of high expression of Axl in NPC was significantly higher than that in NPI(P<0.01).High Axl expression showed no cor-relations with NPC patients,age,gender and M stage,while positively correlated with the clinical stage,T stage and N stage(P<0.05).Axl protein showed a low level in the CNE1cells,but showed a high level in CNE2Z and C666-1cells.TP-0903inhibited cell viability in concentration and time dependent manners.TP-0903at2nmol/L showed significant in-hibitory effects,as evidenced by arresting the cell cycle at G0phase and reducing Axl activity and PCNA expression.CON-CLUSION:High expression of Axl promotes the clinical progress of NPC.TP-0903significantly inhibits the viability of NPC cells,suggesting that Axl may be a valuable target in the NPC treatment.
作者
贾亚楠
李汝佳
王可可
雷洪
哈艳平
王思思
廖晓敏
揭伟
申志华
JIA Ya-nan;LI Ru-jia;WANG Ke-ke;LEI Hong;HA Yan-ping;Wang Sisi;LIAO Xiao-min;JIE Wei;SHEN Zhi-hua(Department of Pathophysiology,Guangdong Medical University,Zhanjiang 524023 , China;Department of Pathology, School of Basic Medical Sciences, Guangdong Medical University ,Zhanjiang 524023 , China.;Pathological Diagnosis and Research Centre,Affiliated Hospital of Guangdong Medical U- niversity,Zhanjiang 524001,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2017年第8期1386-1392,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81402415)
广东医科大学科研基金(No.Z2013004
No.M2013032)
湛江市科技计划项目(No.2013B01077)
作者简介
并列第1作者;通讯作者 Tel: 0759-2388587; E-mail: szh75@126.com
