NM23-H1、DJ1和TIM1在低分化鼻咽鳞癌组织和CNE2细胞株中共同表达  

Over Expression of NM23-H1、DJ1 and TIM1 in Poor Differential Nasopharyngeal Carcinoma Tissues and CNE2 Cell Line

作　　者：冯雪萍[1] 陈主初[1] 肖志强[1] 杨海燕[1] 诸葛勤[1] 朱果[1] 杨轶轩[1] 李明[1] 李萃[1] 李茂玉[1] 李峰[1] 张鹏飞[1] 肖健云[2] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组重点实验室,长沙410008 [2]中南大学湘雅医院耳鼻咽喉科,长沙410008

出　　处：《生物化学与生物物理进展》2005年第4期338-346,共9页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展规划项目(973)(2001CB5102); 国家自然科学基金资助项目(30000028;30240056;30370642);教育部跨世纪优秀人才培养计划基金(教计函[2002]48);湖南省科技厅重大科技专项(02SSY2001-1) ;湖南省卫生厅重点科研项目(Z02-4).~~

摘　　要：鼻咽癌为一种好发于鼻咽项前壁及咽隐窝的头颈肿瘤,98%属低分化鳞癌.鼻咽癌就诊的患者中约75%已有颈淋巴结转移,颅神经受累(Ⅱ～Ⅵ,Ⅸ～Ⅻ)也较易发生.CNE2为一种低分化鼻咽鳞癌细胞系,其生物学行为具有低分化鼻咽鳞癌的一些共同特点.选择恶性程度高的鼻咽癌组织(Ⅳ期,低分化鼻咽鳞癌)和恶性程度高的低分化鼻咽鳞癌细胞株CNE2为研究样本,应用双向凝胶电泳技术获得了6例分辨率较高、重复性较好的低分化鼻咽鳞癌组织和CNE2细胞系双向凝胶电泳图谱,利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术,分析了145个蛋白质斑点(组织66个点,细胞株79个点),共鉴定出了74种蛋白质,其中瘤基因DJ1、转移相关基因NM23-H1和磷酸丙糖激酶异构酶1(TIM1)3种蛋白质,在CNE2细胞系和该6例低分化鼻咽鳞癌组织中均共同表达.并进一步应用RT-PCR法检测低分化鼻咽鳞癌细胞系CNE2和12例低分化鼻咽鳞癌组织三者mRNA的表达:在细胞系CNE2和3例Ⅳ期低分化鼻咽鳞癌病人的组织中三者全表达,3例Ⅲ期及6例Ⅱ期也各有1例为全表达,而在正常对照的6例慢性鼻咽炎组织中未检测到三者的同时表达且有一例为全阴性,NM23-H1、DJ1和TIM1三者的mRNA在Ⅲ、Ⅳ期低分化鼻咽鳞癌组织共同表达,与正常对照组织相比,发现两者有显著性差异(x2=10.214,P<0.005).NM23-H1、DJ1和TIM1三者的共同表达,可能与低分化鼻咽鳞癌的发生发展有关.Nasopharyngeal cancer (NPC) is a type of head and neck cancer and occurs in the uppermost region of the throat situated behind the nasal cavity.98% of them are poor differential nasopharyngeal squamous carcinoma. 75% of NPC were found with enlarged lymph nodes in. the neck, and also usually cranial nerve dysfunction (usually II similar to VI or IX similar to XII). CNE2 is a kind of poor differential nasopharyngeal squamous carcinoma cell line, which existed some vital characteristic of poor differential nasopharyngeal squamous carcinoma. High malignant NPC tissues (in IV -stage, poor differential nasopharyngeal squamous carcinoma) and high malignant poor differential nasopharyngeal squamous carcinoma cell line CNE2 as the test samples were chosed and good 2-DE patterns of the 6 cases nasopharyngeal squamous carcinoma tissues and poor differential pharyngeal were established squamous carcinoma cell line CNE2 proteins with high resolution and good reproducibility using Immobilized pH gradient two-dimensional technology. Total 145 protein spots (66 spots in the tissues and 79 spots in the cell line) were analyzed by MALDI-TOF-MS, and 74 protein spots were identified. 3 proteins of the identified proteins were found to over express in poor differential pharyngeal squamous carcinoma cell line, as well as in 6 cases IV -stage nasopharyngeal squamous carcinoma tissues. Their mRNAs in cell line CNE2 and in 14 cases of human nasopharyngeal squamous carcinoma tissues were tested by RT-PCR. All mRNAs of TIM1, DJ1 and NM23-H1 were expressed both in cell line CNE2 and in tissues (3 cases in IV -stage, 1 cases in III -stage and 1 cases in II -stage, but none of 6 cases in chronic epithelium tissues), and one of 6 cases chronic epithelium tissues did expressed none of TIM1, DJ1 and NM23-H1 mRNAs. mRNAs of nm23-H1, DJ1 and TIM1 in III, IV-stage NPC tissues were expressed simultaneity, which were compared with chronic nasopharyngitis epithelium tissues, were existed the evidently difference (x(2)=10.214, P<0.005). The over expres

关 键 词：低分化鼻咽鳞癌 2-DE MALDI—TOF—MS 蛋白质组 RT—PCR 

分 类 号：R739.6[医药卫生—肿瘤]