| Objective:The purpose of this study was to screen differentially expressed mRNAs in OSCC and normal tissues of Chinese hamster by high-throughput RNA sequencing and real-time qPCR, and to explore the intrinsic mechanism of OSCC development and progression from the perspective of transcriptome and gene expression, and to explore the molecular mechanisms and clinical significance of candidate genes in the invasion and metastasis of tumor.Methods:We successfully builded Chinese hamster’s oral buccal mucosa squamous cell carcinoma experimental animal model induced by 9,10-Dimethyl-1,2-Benzanthracene(DMBA). We obtained the differentially expressed mRNA expression profiles in fresh-frozen OSCC tissue specimens and normal tissue specimens of Chinese hamster by high-throughput RNA sequencing. By using Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses, the functional significance of the differentially expressed genes were analyzed. Meanwhlie we identify the candidate genes such as CXCL1, CXCL2, CXCL3, CCL7, MMP9 by RT-qPCR on OSCC tissue specimens and normal tissue specimens of Chinese hamster.Results:The study showed that, the stages of histological changes in Chinese hamster’s model group have the simple hyperplasia, the epithelial dysplasia, carcinoma in situ and squamous cell carcinomas. We identified 194 significantly differentially expressedmRNA by analyzing Gene expression data of OSCC tissue and normal tissue(|log2Ratio|≥1, Corrected P-value<0.05), of which 66 significantly down-regulated mRNAs and 128 up-regulated mRNAs in OSCC, compared to normal tissue. The differentially expressed mRNAs were involved in 120 biological processes and 8 signal pathways related to OSCC by GO and KEGG analysis, these genes were enriched in multiple Gene Ontology biological processes(e.g., cell chemotaxis, cell migration, cell motility and localization of cell), the mainly significantly signaling pathways activated in OSCC were TNF signaling pathway, chemokine signaling pathway, cytokine-cytokine receptor interaction, ECM-receptor interaction. Compared to normal tissue, the expression of candidate genes CXCL1, CXCL2, CXCL3, CCL7, MMP9 was significantly up-regulated, which was verified by RT-qPCR.Conclusions:In this study, the bioinformatic analysis between OSCC tissue and normal tissue of Chinese hamster by high-throughput RNA sequencing were carried for the first time. we found the abnormal expression of 194 mRNA involved in the normal-cancer process, so8 key signaling pathways including TNF signaling pathway, chemokine signaling pathway, cytokine-cytokine receptor interaction, ECM-receptor interaction were significantly activated. The CXCL1, CXCL2, CXCL3, CCL7, MMP9 genes were over expressed in OSCC and significantly enriched in TNF signaling pathway and chemokine signaling pathway, and may be used as molecular diagnostic markers and therapeutic targats in OSCC, while further research is needed to demonstrate the mechanism of these genes in OSCC progression. |
