Identification of candidate molecular markers of nasopharyngeal carcinoma by microarray analysis of subtracted cDNA libraries constructed by suppression subtractive hybridization
Identification of candidate molecular markers of nasopharyngeal carcinoma by microarray analysis of subtracted cDNA libraries constructed by suppression subtractive hybridization
复制标题
抑制消减杂交构建的消减cDNA文库微阵列分析鉴定鼻咽癌候选分子标志物
DOI：
10.1097/cej.0b013e328305a0e8
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发表时间：
2008-11-01
影响因子：
2.4
通讯作者：
Li, Guiyuan
中科院分区：
文献类型：
--
作者：
Zhou, Yanhong;Zeng, Zhaoyang;Li, Guiyuan
筛选鼻咽癌差异表达基因并筛选候选分子标记。采用抑制性消减杂交技术构建了4个消减cDNA文库，随机挑选约1200个克隆构建cDNA微阵列，分析了19个NPC、3个NPC来源细胞系和10个鼻咽粘膜慢性炎症组织的基因表达谱。我们使用实时定量逆转录聚合酶链反应和原位杂交技术来证实我们的微阵列结果。结果显示，鼻咽癌组织中有37个高表达集落，68个低表达集落。通过对105个鼻咽癌差异表达克隆进行测序，筛选出32个已知基因。在68个NPC低表达集落中，腭、肺和鼻上皮癌（PLC-associated and homo sapien cell division cycle 37 homolog（Saccharomyces cerevisiae）-like 1，CDC 37 L1）基因的频率较高。在37个鼻咽癌高表达克隆中，信号转导和转录激活因子5A基因频率最高，其次是RAS癌基因家族成员和分泌蛋白、酸性、富含半胱氨酸基因。实时定量逆转录-聚合酶链反应和原位杂交技术检测结果显示，鼻咽癌组织中PLR和CDC 37 L1的表达率均低于鼻咽黏膜慢性炎症组（P<0.01）。结果提示，PLC 10和CDC 37 L1基因可能是鼻咽癌的分子标记。首次发现CDC 37 L1与鼻咽癌有密切关系。
To identify differentially expressed genes and scan candidate molecular markers in nasopharyngeal carcinoma (NPC). We constructed four subtracted cDNA libraries using suppression subtractive hybridization technique, then randomly picked about 1200 colonies from the libraries to construct cDNA microarray and analyzed the gene expression profile in 19 NPCs, three NPC-derived cell lines, and 10 chronic inflammation of nasopharyngeal mucosa tissue samples using the cDNA microarray. We used real-time quantitative reverse transcription polymerase chain reaction and in-situ hybridization techniques to confirm our microarray results. The results showed that there were 37 highly expressed colonies and 68 poorly expressed colonies in NPC. Thirty-two known genes were identified by sequencing 105 differentially expressed colonies in NPC. Palate, lung, and nasal epithelium carcinoma (PLUNC)-associated and homo sapien cell division cycle 37 homolog (Saccharomyces cerevisiae)-like 1 (CDC37L1) genes had a higher frequency than others in the 68 poorly expressed colonies in NPC. The frequency of signal transducer and activator of transcription 5A gene was the highest in the 37 highly expressed colonies in NPC; after that were member RAS oncogene family and secreted protein, acidic, cysteine-rich genes. Real-time quantitative reverse transcription-polymerase chain reaction and in-situ hybridization techniques confirmed that the NPC group had a lower frequency of PLUNC and CDC37L1 expression than the groups of chronic inflammation of nasopharyngeal mucosa (P<0.01). The data suggested that PLUNC and CDC37L1 genes might be the putative molecular markers of NPC. For the first time we found that there was a close relationship between CDC37L1 and NPC.