Gene Expression Profile And Molecular Marker Screening Of Allergic Rhinitis In Uygur Nationality In Xinjiang

Objective: through the construction of the Xinjiang Uygur allergic rhinitis mRNA map,whether mRNA is differentially expressed in Xinjiang Uygur patients with allergic rhinitis in the normal nasal mucosa,and selected the significant mRNA,of mRNA in the process of action and possible mechanism of allergic rhinitis in Xinjiang uygur.Methods: 1)RNA-Seq construction and screening of allergic rhinitis in Xinjiang study on gene expression profiles of Xinjiang Uygur,allergic rhinitis and allergic rhinitis patients with 3 cases in each gene expression spectrum of nasal mucosa was detected by RNA-Seq technique,and the differences found by screening.2)analysis of gene expression information of molecular biology in Xinjiang the difference of allergic rhinitis.On screening of differentially expressed genes,the preliminary analysis of differentially expressed genes GO and KEGG enrichment analysis were carried out.3)quantitative PCR validation study of differentially expressed genes in allergic rhinitis in Xinjiang Uygur population.Collected 8 cases of patients with allergic rhinitis in Xinjiang Uygur and 6 cases of patients with allergic rhinitis nasal mucosa CXCL5 gene(chemokine family(IL6),interleukin 6(ROCK2)and related serine / threonine protein kinase(STAT1),signal transduction and transcription activation promoter)study of quantitative PCR validation.Result: 1)RNA electrophoresis showed that 18 S and 28 S two RNA r bands were clear,no genomic DNA contamination.The extracted RNA had good integrity and purity,which indicated that RNA was successfully extracted from nasal mucosa,and the experimental results of mRNAseq were satisfied.In 6 samples were detected 23284 genes,and non allergic rhinitis group specimens to gene expression was detected in 1,2,6 in the sample(at least the expression level of >0 in a biological repeat gene)20541(of which 1132 genes only in non allergic rhinitis,allergic cases express)rhinitis to gene expression was detected in 3,4,5 in the sample(at least the expression level of >0 in a biological repeat gene)19676(of which 267 genes only in allergic rhinitis samples;in addition,the expression of)a total of 20808 genes was detected in specimens or non allergic rhinitis allergic rhinitis samples(>0 gene expression level in the sample,any)19409 genes simultaneously in non allergic rhinitis and allergic rhinitis specimens were detected in the expression(at least in the non allergic rhinitis and allergic rhinitis were specimens Gene expression level in a biological repeat >0.Count>=2 and pvalue<0.01,the difference between the representative signal strength of at least 2 times,this study set the difference of at least 2 times more meaningful,can have 230 different expression genes,97 up-regulated genes and 133 down regulated expression.2)using Gene Spring software,two differentially expressed genes according to the Gene Ontology database on the expression of biological processes involved in gene(biological process),cellular component,molecular function(cellular component)(molecular function)three aspects of functional annotation,found the 402 enrichment of GO function(enrichment the standard for Count>=2 and pvalue<0.01,were selected in the GO function and transcriptional regulation,response and response regulation,signal transduction,signal transduction,cell proliferation,cell proliferation,metabolism and metabolic regulation,receptor binding related differentially expressed genes.According to the analysis of the KEGG database of 230 genes between biological pathways(pathway)analysis: in this study we obtained 18 pathways respectively(Salivary secretion,Calcium signaling pathway saliva)(calcium signaling pathway),Chemokine signaling pathway(chemokine signaling pathway),Gap junction(gap junction),African trypanosomiasis(African trypanosomiasis),GnRH signaling pathway(GnRH signaling pathway),Epithelial cell signaling in Helicobacter pylori infection(Helicobacter pylori infection of epithelial cell signal),Long-term potentiation(LTP),Sphingolipid(metabolism,Graft-versus-host disease(sphingolipid metabolism),graft-versus-host disease)N-Glycan biosynthesis(N-,Cytokine-cytok polysaccharide biosynthesis)Ine receptor interaction(cytokine receptor interaction),Malaria(Nve Ji),Osteoclast differentiation(osteoclast differentiation),Arginine and proline metabolism(arginine and proline metabolism),Circadian rhythm(circadian rhythm),Rheumatoid arthritis(rheumatoid arthritis),Hepatitis C(hepatitis C).Pay attention to the 3 pathways were signaling pathway(calcium signaling pathway),Chemokine signaling pathway(chemokine signaling pathway),Cytokine-cytokine receptor interaction(cytokine receptor interaction),the expression level of 17 genes corresponding to the change of more than 2 times,which can be used for subsequent experiments.Up and down the most significant genes further analysis of nasal mucosa in Uygur patients with allergic rhinitis and non allergic rhinitis 23284 gene expression profile of RNA-Seq detection,the results showed that the expression of allergic rhinitis in the spectrum of 230 differentially expressed genes,including 97 up-regulated genes and 133 down regulated genes,the expression level of 17 genes corresponding to the more than 2 times change,we found by analyzing the-down regulated genes ADCY8,CXCL5,IL6,CSF3,CXCL3,NOS2,CXCL2,CXCL1,SPHK2,ITPR1,F2 R,up regulated genes STAT1,EGFR,ROCK2,P2RX7,GNAQ,CXCL9.3)the expression of STAT1 and ROCK2 was down regulated by PCR-qRT,and the expression of IL6 and CXCL5 was upward trend;the results were consistent with the RNA-seq prediction.Conclusion: there is a significant difference between the expression of RNA m and normal nasal mucosa in the nasal mucosa of allergic rhinitis patients in Xinjiang Uygur Autonomous Region.The expression level of mRNA is not equal to the protein(protein and its protein level is representative of the final result),gene expression depends on the encoding protein function.In short,there should be a comprehensive,reasonable and objective understanding and explanation of the differences in mRNA levels.This result requires large samples,multiple levels of clinical validation.