Search for oral cancer specific genes and molecular cloning using in-house cDNA microaray

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cDNA library was constructed from surgical specimens of oral squamous cell carcinoma(SCC) and salivary gland tumors(SGT) using an oligo-eapping method. The 5 '-end nucleotide sequences of the cDNA clones were determined. Overlapping clones were excluded, and 2,201 independent clones were selected and used for microarray production. Compared to the public nucleotide sequence database, 61% of our cDNA clones were full length. mRNAs obtained from squamous cell carcinomas(SCC) and adenoid cystic carcinomas(ACC) were analyzed using this microarray system. Microarray detected up-regulated expression of many genes, which expressed twice or more in tumors man in normal tissue. We could pick up the full length whole gene, which was focused on by microarray analysis, and were directly and immediately able to examine the gene function using the full length clone.Representative full length clones with differential up-/down-regulated expression in SCC were Interleukin 1 β, Centaurin gamma 2, Adenyl … More yl cyclase-associated protein, Rh type C glycoprotein, Spermine syntase, Copine 1, Cytokeratin 13, Paired-like homeodomain transcription factor 1, Hypothetical protein FLJ20150, Small proline-rich protein, Cartilage link protein, G-protein alpha 15, Interleukin receptor antagonist, Transglutamin and so on, whereas the genes differentially expressed in SGTs were decay accelerating factor for complement (CD55), Inducible signaling pathway protein 2, Insulin-like growth factor binding protein 3, Fos-like antigen 1,Human vimentin gene, Immunogloblin heavy constant mu, T cell receptor beta chain, Dehydrogenase/reductase member 2,18S rRNA gene and et. The fgene function -nalysis by edycsone-promoter system and immunohistochemical staining of clinical samples, revealed several important bio-markers. Clorf 10 gene is a tumor suppressor, which stops cell cycle, Rab la is a candidate for a predictor of carcinogenesis, maspin is a molecular marker specific to ACC, and stathmin is a molecular marker specific to malignancy. Less