Identification of candidate molecular markers of nasopharyngeal carcinoma by microarray analysis of subtracted cDNA libraries constructed by suppression subtractive hybridization

Abstract

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.