反义细胞角蛋白13基因对人鼻咽癌HNE1细胞移植瘤放疗敏感性的影响  

Effect of antisense CK13 genes on human nasopharyngeal carcinoma HNEl cell transplantation tumor

作　　者：苗春雨[1] 余宏[1] 

机构地区：[1]昆明医科大学第四附属医院耳鼻咽喉科,云南昆明650000

出　　处：《中国耳鼻咽喉头颈外科》2016年第5期253-258,共6页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：云南省科技厅昆明医科大联合专项资金资助(2012FB084)

摘　　要：目的观察反义细胞角蛋白13(cytokeratin 13,CKl3)基因对鼻咽癌人嗜中性细胞弹性蛋白酶1(human neutrophil elastase,HNE1)细胞移植瘤放疗敏感性的影响。方法将HNE1细胞株分为A组(未经处理)、B组(转染慢病毒空载体),C组(转染慢病毒反义CK13a)和D组(转染慢病毒反义CK13b)共4组建立相应动物模型,放疗后采用流式细胞仪、免疫组化、PCR、Tunel法及Westernblotting检测。结果细胞周期检测发现转染慢病毒质粒的C组和D组在放疗后与对照组比较G2/M期阻滞时间显著延长;免疫组化结果显示C组和D组移植瘤中CK13表达下降;Tunel法检测细胞凋亡坏死率发现C组和D组细胞凋亡率明显降低;Western blotting检测发现caspase-3凋亡标志物下降。PCR检测发现CDC25mRNA水平明显降低。结论反义CK13基因通过调控细胞周期和凋亡可降低鼻咽癌HNE1移植瘤的放疗敏感性,并与caspase-3凋亡途径和CDC25信号通路相关。OBJECTIVE The influnence of observation on the antisense cytokeratin 13（CK13） gene in nasopharyngeal carcinoma HNEl cell transplantation tumor radiation sensitivity.METHODS HNEl cell lines can be divided into control group：the control group（HNEl cell）and lentivirus（transfection slow virus empty carrier） group and experimental group：HNEl-anti-CK13a（transfection antisense CK13 a slow virus） and HNEl-anti-CK13b（transfection antisense slow virus CK13b） four groups,set up a corresponding animal model,after radiotherapy by flow cytometry,immunohistochemistry,PCR,Tunel method and Westernblotting detection.RESULTS The cell cyclede tection of plasmid transfection slow virus group compared with control group after radiotherapy G2/M phase of the block were significantly prolonged;Immunohistochemical results showed emigration tumor CK13 expression decreased in the experimental group;Tunel method to detect apoptosis necrosis rate found that the experimental group,apoptosis rate significantly decreased;Western blotting detection caspase 3 apoptosis markers.PCR to detect CDC25 mRNA level decreased obviously.CONCLUSION Antisense CK13 gene by regulating the cell cycle and apoptosis can reduce HNEl transplantation tumor radiotherapy of nasopharyngeal carcinoma（NPC） sensitivity,and with the caspase 3 apoptotic pathways and CDC25 signaling pathways.

关 键 词：鼻咽肿瘤 细胞凋亡 细胞周期 反义细胞角蛋白13 

分 类 号：R739.63[医药卫生—肿瘤]