Screening And Analysis Of Tumor Associated Genes And LncRNAs In Chicken MD

Marek’s disease virus（MDV）is an important pathogen of avian diseases.The pathogenic type MDV-1 can cause chicken infectious disease,which is characterized by severe immunosuppression and T lymphocyte tumor.Marek’s disease（MD）tumor induced by MDV is an ideal animal model for the study and treatment of viral tumors.In many studies,it is found that the Long non-coding RNAs（lncRNAs）has abnormal expression,up or down regulation of IncRNAs occurs at various stages of tumorigenesis,it may promote or inhibit the formation and development of tumor.In this study we used MD spleen tumor and normal spleen tissue RNA-Seq obtained by gene expression database,database,analyzing the difference signal pathway database and gene interaction database by analyzing the differentially expressed genes associated with cancer screening,gene expression experiments and one of the important,to explore its expression form and in the process of tumor MD function.The analysis results show that:（1）There are 499 differentially expressed genes were annotated by 256 signaling pathways,6 signaling pathways associated with the tumor showed a significant difference（P<0.05）,tumor necrosis factor（TNF signaling pathway）signaling pathway is associated with tumorigenesis was highest（P<0.05）.（2）In MD tumor tissue,the level of mRNA,SOCS1 and SOCS3 gene expression of suppressor of cytokine signaling up-regulated,at the core position in gene regulatory networks,the downstream inflammatory factors to inhibit tumorigenesis,influence the process;co expression of SOCS1 and SOCS3 genes were also up-regulated the expression of NF-κB and signal transducer and activator of transcription factor STAT1 and STAT3 genes,the formation of the two kinds of regulatory factors can promote tumor.Based on the screened genes of qPCR results showed that:compared with the control group,MDV infection group 5 genes screened showed up-regulated expression,and the RNA-Seq prediction analysis results,RNA-Seq results reflect the MD tumor gene expression in the process of difference.Secondly,with the new IncRNAs database as the data source analysis tools,used to calculate potential（Coding Potential encoding Calculator,CPC）,assessment tools（Coding Potential Assessment Tool encoding potential,CPAT）,sequence alignment tool（blastx）and Ensembl BLAST/BLAT and other online analysis tools,further analysis of cross validation for protein encoding ability of the new the entries in the IncRNAs database,from screening with MD tumor related IncRNAs.The analysis results obtained 6425 new IncRNAs with lower coding ability.Among them,the six IncRNAs selected by TCONS₀0025968,TCONS₀0032120,TCONS₀0036544,TCONS₀0009801,TCONS₀0009802 and TCONS₀0009803 may be related to the expression of tumor suppressor p53 gene.Application of chicken MD animal model of MDV infection in first,twenty-first,thirty-fifth and 42 days of age,to detect the transcription level of p53 gene in chicken spleen tissue samples and TCONS₀0009803,TCONS₀0009801 and TCONS₀0009802 three lncRNAs,with the same age normal chicken spleen comparison results show that:（1）After MDV infection,the transcriptional level of p53 gene and TCONS₀0009801 showed a consistent change trend during 1～35 days of age increased,decreased at 42 days of age,MDV showed that in the early stage of the invasion,the two have synergistic effect.With the prolongation of MDV in host body,the expression of lncRNA-TCONS₀0009801 and p53 is down regulated,which may be related to apoptosis,dysfunction and immortalization of cells.（2）After MDV infection increases with age,TCONS₀0009802 and TCONS₀009803 expression level was increased gradually,the difference and 1 day of age at the age of 21 days was not significant（P>0.05）,and at the age of 35 days and 42 days of age were significant differences（P<0.01）.In the spleen of the normal chicken,the trend of first rise and then decrease,and the two IncRNAs did not change significantly with the increase of the age of the chicken（P>0.05）.After MDV infection,the transcription of lncRNA-TCONS₀0009801 and p53 gene of chicken spleen tissue expression change trend was consistent raised after the first cut;but TCONS₀0009802 and TCONS₀0009803 increased with the increasing age of chicken p53 gene transcription level,while over the same period the transcription level in the 1～35 age first increases,and then in the 42 day of the downward trend is not consistent three,the expression of IncRNAs on transcription of p53 gene may play different roles,its influence on the function of p53 still need further discussion.The expression levels of p53 gene and three lncRNAs were significantly different between the experimental group and the control group at the other three days except the 1 day old age（P<0.01）,At the age of 42 days,the transcription levels of the three lncRNAs above were consistent with those predicted by RNA-Seq analysis,indicating that the RNA-Seq results truly reflect the transcription of lncRNAs in the course of MD tumorigenesis.The MDV induced tumor tissue transcriptome information and the differential expression of related genes in this study provide a valuable reference for the study of the pathogenesis of viral tumor.