3例喉咽癌组织与相邻正常黏膜的基因表达谱差异分析  

Differentially expressed genes in 3 cases of hypopharyngeal carcinoma and normal tissues by cDNA microarray

作　　者：吕梅[1] 董频[2] 佘翠萍[3] 杜翠萍[1] 李勇[1] 徐尔东[1] 

机构地区：[1]大连医科大学附属第一医院耳鼻咽喉科,辽宁大连116011 [2]上海交通大学附属第一人民医院耳鼻咽喉-头颈外科,上海200080 [3]大连市中心医院耳鼻咽喉-头颈外科,辽宁大连116033

出　　处：《山东大学耳鼻喉眼学报》2010年第5期5-9,28,共6页Journal of Otolaryngology and Ophthalmology of Shandong University

摘　　要：目的研究喉咽癌组织与相邻正常黏膜的基因表达谱差异,从基因组水平初步探讨喉咽癌易侵袭和转移的机制。方法 3例经病理证实为鳞状细胞癌的喉咽癌标本和邻近的正常黏膜组织标本,提取细胞总RNA,反转录为Cy3标记的癌旁正常喉黏膜cDNA,及Cy5标记的喉咽癌cDNA,与共有人类癌症相关基因886条的基因表达谱芯片Human Cancer CHIP Ver.4.0进行杂交,杂交芯片用双通道激光扫描仪进行扫描记录,利用ImaGene分析软件将信号强度数值化。基因显著性表达的判定标准为:每个基因的喉咽癌组织和癌旁正常组织的平均灰度比值R(Cy5/Cy3)相差2倍或2倍以上,则认为差异有统计学意义。结果在3例样本中重复表达且相差2倍以上的基因共51个,喉咽癌组织表达上调者39个,表达下调者12个。本实验发现显著差异的新基因4个,这些基因序列已被测出,但其功能却未知。其中表达差异非常显著(平均相差5倍以上)的主要包括,上调基因:基质金属蛋白酶-1(MMP-1),整合素β4(Integrinβ4),Bcl-2,CXCL13,黑色素瘤特异性抗原(PRAME);下调基因:早期生长反应基因-1(early growth respons gene-1,Egr-1),MRP1/CD9。结论基因芯片技术可以用来筛选在喉咽癌肿瘤细胞增殖、侵袭、转移过程中发挥重要作用的多种基因,为进一步深入研究提供靶点。Objective To detect differentially expressed genes in hypopharyngeal carcinoma and adjacent normal laryngeal tissues with cDNA microarray.Methods Three samples of hypopharyngeal carcinoma,pathologically confirmed as squamous cell carcinoma,were tested.Total RNAs were extracted and inversely transcribed to cDNAs.The cDNAs of hypopharyngeal carcinoma were marked by Cy5 and the cDNAs of normal laryngeal tissues were marked by Cy3.The cDNA probes were then hybridized to DNA microarrays and scanned for fluorescent signals.Differential expression between the two tissues was produced by ImaGene software.The average ratio of gray scale,if equal or more than 2 times,was considered significant.Results There were 51 genes significantly expressed in 3 cases.Among them,39 genes were significantly higher,but 12 genes were significantly lower in hypopharyngeal carcinoma than normal tisscle.4 new genes were identified.Some highly expressed genes（more than 5 fold difference） included： 1） Genes of upregulation,such as MMP-1,Integrinβ4,Bcl-2,CXCL13 and PRAME;and 2） Genes of downregulation,such as Egr-1 and MRP1/ CD9.Conclusion Many genes,which play important roles in the proliferation,invasion,and metastasis of hypopharyngeal carcinoma,can be picked out by cDNA microarray;These genes will serve as targets for further research.

关 键 词：喉咽癌 鳞状细胞癌 基因表达 基因芯片 

分 类 号：R739.65[医药卫生—肿瘤]