Role of Chemokine and TNF signaling pathway in oral squamous cell carcinoma: A RNA deep sequencing analysis of oral buccal mucosa squamous carcinoma model of Chinese hamster

Abstract

16Oral cancer is one of the most common cancers in the world, meanwhile, differentially 17 expressed genes are thought to regulate the development and progression of oral 18 squamous cell carcinomas (OSCC). In this study we screened RNA transcripts from the 19 oral buccal mucosa of healthy male Chinese hamster, divided into 3 groups: a control 20 group with no disposal, a solvent control group coated with acetone solvent, and an 21 experimental group coated with 0.5% DMBA acetone solution by high-throughput 22 RNA sequencing. Tophat and Bowtie were used to align the high-quality reads into 23 transcripts, DEseq was used to analysis the expression of differential gene. Then, the 24 Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and 25 Genomes (KEGG) pathway enrichment analyses were conducted. The chemokine and 26 TNF signaling pathway were differentially expression and the mRNA expression of 27 CXCL1 、 CXCL2 、 CXCL3 、 CCL7 、 MMP9, monitored by qRT-PCR, increased 28 remarkably in the cancer group and coincided with the result of RNA-Sequencing.29 Meanwhile, the CXCL1, CXCL2, CXCL3, and CCL7 are significantly enriched in the 30 chemokine signaling pathway, and CXCL1, CXCL2, CXCL3, and MMP9 are 31 significantly enriched in the tumor necrosis factor (TNF) signaling pathway. The 32 differentially expression of the chemokine and TNF signaling pathway was a response 33 to the invasion of the organism immune system due to oral buccal mucosa squamous 34 carcinoma. All the findings provided novel insights for further molecular researches of 35 oral cancer. 36 Keywords: oral squamous cell carcinoma (OSCC), Chinese hamster, high-throughput 37 sequencing, differentially expressed genes(DEGs) 38 Introduction 39 Oral cancer is one of the most frequent solid cancers worldwide, and oral squamous 40 cell carcinoma (OSCC) constitutes around 90% of oral cancers (Siegel et al., 2014). It 41 is highly invasive and metastatic at the advanced stage, and presents a substantial threat 42 to human health. Meanwhile, there are 145000 deaths of OSCC in the world (1.8% of 43 all cancer deaths in the world) annually; including 77% of the burden is in developing 44 countries (Ferlay et al., 2015). Moreover, the incidence rate of OSCC is increasing, 45 especially in younger people. Furthermore, OSCC has a very poor prognosis due to its 46 invasive nature, the survival rate of patients with OSCC has not improved despite the 47 improvements and innovations in diagnostic techniques and treatments (Shlok et al., 48 2010). However, clinical samples are pretty difficult to obtain and the less number of 49 clinical samples meeting the experimental requirements is a major problem in oral 50 cancer research, which seriously restricts the development of research on the 51 mechanism of OSCC. Therefore, there is an urgent need for a better animal model of 52 human OSCC lesions to help us better understand the pathogenesis of oral cancer. 53 The hamster cheek pouch is the most relevant known animal system that closely 54 related to ...