Whole-transcriptome Sequencing And Differential Expression Of Long Noncoding RNA In Renal Clear Cell Carcinoma

Objective:The whole RNA sequencing was used to draw the gene mutation map of specific renal clear cell carcinoma,to verify the known possible mutations in the occurrence and development of renal clear cell carcinoma,to find unknown non coding RNA affecting the tumor cell microenvironment of renal clear cell carcinoma,and to explore the important signal pathways involved in the mutated genes in renal clear cell carcinoma.Furthermore,the differential expression of mRNA and lncrna at transcriptome level was explored by whole transcriptome sequencing.The pathway enrichment analysis of mRNA and lncrna with significant difference in expression was performed by proteomic analysis method to find out the long non coding RNA involved in the development,growth and tumor thrombus formation of renal clear cell carcinoma.Method:Five cases of renal clear cell carcinoma and their matched normal tissues were collected.One patient had vascular tumor thrombus formation,and the tumor thrombus tissue and normal tissue adj acent to tumor thrombus were collected at the same time.Six pairs(12 cases)of samples were collected and all the coding RNA and non coding RNA were extracted to construct the library.The full transcriptome gene sequence was obtained by the second generation high-throughput sequencing technology.The sequencing data were filtered,compared and assembled,and the coding ability was predicted.The differential expression results of mRNA and incrna were obtained by differential analysis software,and lncrna/mRNA with significant differential expression was obtained.Go/KEGG enrichment analysis was carried out for the differential genes and differential target genes,so as to find the possible enrichment of biological signal pathways.The distribution of differentially expressed genes and the enrichment of pathways at transcriptome level between patients with tumor thrombus formation and those without tumor thrombus formation were compared to find the mutation target genes that may affect the formation of tumor thrombus.Results:1.Through full transcriptional sequencing,23232 differentially expressed mRNA were identified in 5 cases of renal clear cell carcinoma,5680 were significantly different,and 60648 lncmas were identified,12624 of which had significant difference.In a case of clear cell renal cell carcinoma with tumor thrombus formation,22941 differentially expressed mRNA were identified,7608 were significantly different,and 55251 lncrnas were identified,with 15387 having significant difference.73.7%of renal clear cell carcinoma without tumor thrombus had significantly different expression of lncrna than 60%of clear cell carcinoma with tumor thrombus.83,8%of renal clear cell carcinoma without tumor thrombus had significantly different expression of lncrna,and 71.2%of renal clear cell carcinoma tumor thrombus had no overlap2.Five patients with renal clear cell carcinoma(RCC)were enrolled in this Goterm and KEGG pathway enrichment analysis.There were 2355 differentially expressed Goterms,invloving 1884 biological functions,104 cell components and53 molecular functions together with 279 signal pathways.After the prediction of target genes for differential Inrnas,25 goterms were newly enriched,including 16 biological processes,4 cell components,5 molecular functions,and 102 signal pathways were enriched by KEGG enrichment analysis,including PI3K Akt signaling pathway,rapl signaling pathway,ubiquitin mediated proteolysis,snare interaction of vesicle transport,etc.PI3K Akt signaling pathway was the most significant enrichment pathway,and more than 100 lncrnas were significantly enriched in this pathway.Conclusion:1.There are significant differences in genomic and transcriptomic characteristics between RCC with thrombus formation and RCC without tumor thrombus.It is suggested that the molecular mechanism of tumor thrombus formation of renal clear cell renal carcinoma may be different from that of simple renal carcinoma,and the difference in lncma level is more obvious than that in mRNA level,suggesting that lncma may endow some of the tumor cells by regulating the microenvironment of tumor cells.The higher activity of tumor tissue can promote the formation of tumor thrombus2.This study found a large number of novel lncrnas,which were enriched in a large number of biological signal pathways in proteomic analysis.The most significant enrichment pathway was PI3K Akt signaling pathway,and more than 100 lncrnas were significantly enriched in this pathway;suggesting that these unknown lncmas may further regulate the occurrence of tumor by participating in the regulation process of PI3K/Akt/mTOR pathway.