Screen And Bio-informatics Analysis Of Key Disease Genes For Precancerous Lesions Of Oral Buccal Mucosa Induced By DMBA In Golden Hamsters

BackgroundOral cancer accounts for approximately 2% of systemic malignant tumors, of which 90% are squamous cell carcinomas. Buccal cancer is one of the most common oral cancers. Oral cancer is a complex, multiphase, multi-step pathological process involving multiple genetic changes, where a number of genes play various roles at different stages of cancer development. The present study also confirmed carcinogenesis of oral mucosa is a continuous process of dynamic change (normal oral mucosa epithelium→epithelial dysplasia (precancerous lesions)→oral squamous cell carcinoma). So, if effective intervention to prevent the occurrence of precancerous lesions at the molecular level is achieved, the early and effective prevention of oral cancer may also be achieved.ObjectiveAgilent rat whole genome microarray and biological information analysis were used to screen for key disease genes in the transformation of normal buccal mucosa to precancerous lesions in golden hamsters.MethodsHalf a percent of 7,12-Dimethylbenz(a)-anthracene (DMBA) acetone solution was used to establish a model of precancerous lesions in oral buccal mucosa in golden hamsters. Total RNA from normal mucosal tissue and precancerous lesions was extracted to synthesize cRNA single-labeled with Cy3 fluorescent marker, which was then hybridized with the Agilent rat cDNA whole genome microarray containing 41,000 genes/EST sequences. Ratio≥2 and Ratio≤0.5 were the threshold values for screening differentially expressed genes. Screening of differentially expressed genes were performed with functional classification analysis according to Gene Ontology (GO) and pathway analysis (P < 0.05). RT-PCR was used to verify partially differentially expressed genes.ResultsIn the transformation of normal buccal mucosa to precancerous lesions in golden hamster, a total of 1331 genes were expressed differentially, including 1278 known genes, 53 unknown genes, 747 upregulated genes, and 584 downregulated genes. GO analysis showed that these differentially expressed genes can be divided into 8 functional groups such as genes regulating cell physiological processes and genes relating to cell structure. Pathway analysis revealed a total of 14 gene interaction pathways that significantly associated with the 1278 known differentially expressed genes (P < 0.05). Only 21 genes in the 1278 differentially expressed genes were enriched in the 14 pathways. RT-PCR results for Atp6v0d2 upregulation and Sfrp2 downregulation were consistent with the results of the microarray.ConclusionThe occurrence of precancerous lesions in the oral buccal mucosa of golden hamsters was caused by a number of genetic changes that resulted in changes to their respective pathways. Key candidate genes for the formation of precancerous lesions in oral buccal mucosa included Cyp2b13, Orc1L, casp8, CCL5, CXCL12, CCL20, Serping1, P518/Qrfp, F5, TFPI, Vcam1, Fn1, Angpt2, Lcp2, Cxadr, Lyn, Hck, Btk, RGD1564385/fes, Vav1, and IL5ra. Inhibitors that can regulate these genes or pathways effectively may improve treatment methods and chemoprophylaxis of precancerous lesions.