鼻咽癌T3-4N0期患者免疫相关基因的表达  

Gene Expression Profile of Immune Cells from Patients with Nasopharyngeal Carcinoma of Stage T3-4N0

作　　者：肖锡宾[1] 麦海强[2] 曹素梅[2] 张颖[1] 孙蕊[2] 洪明晃[2] 

机构地区：[1]中山大学肿瘤防治中心,实验研究部,广东广州510060 [2]中山大学肿瘤防治中心鼻咽癌科,广东广州510060

出　　处：《癌症》2005年第2期135-140,共6页Chinese Journal of Cancer

基　　金：广东省医学重点专科基金(No.2003-13)~~

摘　　要：背景与目的鼻咽癌患者发生颈部区域淋巴结转移是比较普遍的现象,但鼻咽癌T34N0期患者尽管肿瘤生长已超出鼻咽腔却不表现区域淋巴结转移的特征。为了探讨与这种现象有关的免疫相关因素,本研究对鼻咽癌T3鄄4N0期患者的外周血免疫细胞基因表达进行了研究。方法分离28例鼻咽癌T3-4N0期治疗前患者、例鼻咽癌28T1-2N2鄄3期治疗前患者和56例建康人的外周血淋巴细胞。分别提取这三组外周血免疫细胞的mRNA。以各组的mRNA为模板,反转录制备标有Cy5或Cy3荧光素的cDNA探针,将来自鼻咽癌样品的Cy5cDNA探针分别与来自健康人的Cy3-cDNA探针混合后,分为T1鄄2N2鄄3与T3鄄4N0组分别在480点的免疫相关基因芯片上进行杂交。将Cy5/Cy3<0.5定为鼻咽癌患者免疫细胞中的下调基因,Cy5/Cy3>2.0定为上调基因。以T1-2N2-3与T3-4N0患者免疫细胞的RNA为模板,用实时定量RT鄄PCR方法对2组芯片中Cy5/Cy3值>4.0(4条)和<0.25(3条)基因的表达进行测定。结果在2组基因芯片实验中,共有157条基因的Cy5/Cy3比值有明显的差异。其中106条基因的Cy5/Cy3值下调,条基因的51Cy5/Cy3值上调。实时定量RT鄄PCR检查显示,在来自T3鄄4N0期患者的样品中编码CD86、CD69、NFKB的基因的表达量比来自T1鄄2N2鄄3期患者的样品高。结论鼻咽癌T1鄄2N2鄄3与T3鄄4N0期群?BACKGROUND & OBJECTIVE: Nasopharyngeal carcinoma (NPC) patients commonly have neck regional lymph node metastases. However, patients with NPC of stage T3-4N0 have no lymph node metastases, even the tumors grow over the nasopharyngeal cavities. This study was to explore immune factors related to such a situation by detecting gene expression profile of immune cells from patients with NPC of stage T3-4N0. METHODS: Peripheral blood lymphocytes were isolated from 28 patients with NPC of stage T1-2N2-3, 28 patients with NPC of stage T3-4N0 before treatment, and 56 healthy donors. mRNA of the isolated lymphocytes was extracted, and reverse-transcribed into cDNA probes labeled with Cy5 or Cy3 fluorescent dyes. The Cy5-cDNA probes of T1-2N2-3 group and T3-4N0 group were separately mixed with Cy3-cDNA probes of healthy group, and hybridized with 480 dots of immune genechips. Genes were considered to be differently expressed in NPC patients if the Cy5/Cy3 was >2.0 (up-regulated), or <0.5 (down-regulated). The genes with Cy5/Cy3 of <0.25 or >4.0 were amplified by real time quantitative reverse transcription-polymerase chain reaction (RT-PCR) with RNA templates of T1-2N2-3 group, and T3-4N0 group, respectively. RESULTS: Of 157 genes showed different expressions, 106 down-regulated, 51 up-regulated. Real time quantitative RT-PCR revealed that among the 4 genes with Cy5/Cy3 of >4.0, and the 3 genes with Cy5/Cy3 of <0.25, expressions of the genes encoding CD86, CD69, NFKB were higher in sample from T3-4N0 group than in sample from T1-2N2-3 group. CONCLUSIONS: More genes are down-regulated in immune cells from patients with NPC of stage T1-2N2-3 or T3-4N0 than those from healthy donors. Whether the different expressions of genes encoding CD86, CD69, and NFKB in immune cells of patients with NPC of stage T1-2N2-3 and stage T3-4N0 related to neck regional lymph node metastasis is worthy of further study.

关 键 词：基因芯片 鼻咽肿瘤 免疫细胞 

分 类 号：R739.63[医药卫生—肿瘤] R730.2[医药卫生—临床医学]