Expression And Analysis Of LncRNAs In Papillary Thyroid Carcinoma With Lateral Lymph Node Metastasis

Objectives:Taking the papillary thyroid carcinoma（PTC）patients with lateral lymph node metastasis（LLNM）as the research object,we use the technology of long non-coding RNA（lncRNA）sequencing and bioinformatics analysis to screen the differentially expressed lncRNAs in cancerous tissue and para-cancerous tissue,to explore their potential functions in the occurrence of LLNM in PTC.It may provide theoretical basis for early diagnosis of the disease.Methods:Following the strict inclusion and exclusion criteria,three PTC patients with LLNM were collected.Samples of the cancerous tissue,para-cancerous tissue are collected to detect the differentially expressed lncRNAs by lncRNA sequencing.Then we predict the target genes of lncRNA with significant differential expression,and conduct the GO enrichment analysis and the KEGG enrichment analysis on these target genes.Measure the length and number of branches of lymphatic vessels after the expression of the highest differential expression lncRNA was knock downed.In order to test and verify the effect of the highest differential expression lncRNA on lymph-angiogenesis.Results:1.There are a quantity of differentially expressed lncRNAs in the cancerous tissue and para-cancerous tissue of PTC patients with LLNM.There are 8589 differentially expressed lncRNAs,of them,4782 up-regulated and 3807 down-regulated.2.Set the standard as FDR<0.05 and|log₂FC|≥1 to select lncRNAs with significant differential expression.It was found that there were 1221 lncRNAs with significantly up-regulated expression and 1511 with significantly down-regulated expression.Lnc RNA SLC26A4-AS1 is the most significantly differentially expressed lncRNA,followed by the lncRNA H19.3.Predict the target genes of those lncRNAs which are expressed differentially.Among them,there are 393 differentially expressed lncRNAs have the target genes,include lncRNA SLC26A4-AS1.4.The background expression of lncRNA SLC26A4-AS1 in human normal thyroid cell line Nthy ori3-1 and thyroid cancer cell lines TPC-1,SW579,and B-CPAP was detected by polymerase chain reaction（qPCR）.The results showed that the expression of lncRNA SLC26A4-AS1 was most obviously significant in TPC-1.5.After knocking down the expression of lncRNASLC26A4-AS1 in TPC-1,the length and the number of branches of intact lymphatic vessels significantly decreased,with a statistically significant difference compared to human normal thyroid cell line Nthyori3-1（P<0.05）.Conclusions:1.There are a quantity of differentially expressed lncRNAs in the cancerous tissue and para-cancerous tissue of PTC with LLNM.2.The expression of lncRNASLC26A4-AS1 in PTC tissue is significantly increased, which may promote LLNM in PTC by promoting lymph-angiogenesis.