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[Identification of differentially expressed genes in primary cultured nasopharyngeal carcinoma cells by cDNA
Rui-Ping Li1, Jian-Yong Shao, Ling Deng
1State Key Laboratory of Oncology in South China and Department of Pathology, Cancer Center, Sun Yat-sen University, Guangzhou 510060, China. wyf12345678@263.net
Objective:
To analyze the global gene expression profile of primary cultured nasopharyngeal carcinoma (NPC) cells using cDNA microarray techniques to screen new candidate genes related to the occurrence and progression of NPC.
Methods:
A NPC cell line C666 and primary cultured NPC cells from biopsy specimens in 5 cases were analyzed with microarray techniques in comparison with 3 normal nasopharyngeal epithelial (NPE) biopsy specimens. Several differentially expressed genes identified from the microarray results were verified by fluorescence real-time PCR (FQ-PCR) and immunohistochemistry (IHC).
Results:
Primary cultured cells of both NPC and NPE were verified by cytokeratin IHC, EBER1 in situ hybridization and EBV-DNA real-time PCR. Compared with NPE cells, a total of 493 genes in at least 4/6 of the samples were identified to be differentially expressed in the primary cultured NPC cells, including 264 up-regulated and 229 down-regulated ones. Several differentially expressed genes according to the microarray results were confirmed by real-time PCR and IHC.
Conclusion:
cDNA microarray technique provides an effective and accurate means for global gene expression profiling of primary cultured NPC cells to screen the differentially expressed genes, which may serve as an important basis for studying the mechanism, classification and diagnosis of NPC at the molecular level.
