Study On The Expression Of Long Non-coding RNA In Thyroid Cells Of Patients With Autoimmune Thyroiditis

Purpose: Recently,the prevalence of clinically dominant autoimmune thyroid disease(AITD)is 2-3%,while the prevalence of thyroid autoantibodies in the general population exceeds 15%.AITD can increase the risk of non-thyroid diseases such as cardiovascular disease,cancer,and adverse pregnancy outcomes,thus causing widespread attention.The thyroid is an important organ of the human body,and the destruction of structure and the progressive loss of function make the process of Hashimoto's thyroiditis(HT)complicated.Although we have discovered Hashimoto's thyroiditis a century ago,its pathogenesis is still unclear.At present,it is believed that the interaction of genetic susceptibility,environmental factors and immune dysregulation factors have an impact on the occurrence of the disease.Long non-coding RNA(lnc RNA)can regulate the immune response,including T cell differentiation,dendritic cell function and cytokine production,and therefore play a role in autoimmune diseases in a cell type-specific manner.lnc RNA participates in the immune response at the level of transcription,post-transcription and translation without changing the genetic material to maintain immune homeostasis.In this study,high-throughput sequencing and subsequent verification of gene and transcriptome m RNA and lnc RNA were performed by collecting RNA from thyroid cells,thus,the expression of long-chain non-coding RNA in primary thyroid cells was analyzed.Method: First,16 cases of Hashimoto's thyroiditis(HT)and 16 cases of normal thyroid tissue(NC)were collected in the operating room,then digested and separated,and cultured for primary thyroid cells,subsequently subjected to immunofluorescence identification.Secondly,the Illumina Hi Seq platform was used to sequence and the Sequencing length was 50bpx200 m.Thirdly,we collected 30 cases of Hashimoto's thyroiditis(HT)and 30 cases of normal thyroid tissue(NC),cultured thyroid cells and then collected RNA,and verified the expression of differential lnc RNA in primary thyroid cells using Real-time PCR Happening.Results: 1.In this high-throughput sequencing,the m RNA,LncRNA genes and transcripts in the thyroid cells of the HT group and the NC group were compared,and it was found that the number of up-regulated differential lnc RNAs genes is 2573 in the HT group and the number of down-regulated differential lnc RNAs genes is 15778.2.After selecting 37 lnc RNAs for Real-Time PCR verification,there were 6 differential lnc RNAs(P <0.05).Among them,the lnc RNAs of NONHSAT185097.1,NONHSAT152327.1,NONHSAT183385.1,NONHSAT213829.1,NONHSAT245695.1were all related to thyroid antibodies.Conclusions: 1.Through KEGG analysis of the differential genes between groups,it is found that the differential lnc RNAs are mainly enriched in autoimmune thyroid disease,systemic lupus erythematosus,rheumatoid arthritis,primary immune deficiency,and other diseases,as well as B cell receptors Signaling pathway,NF-?B signaling pathway,natural killer cell-mediated cytotoxicity,etc.2.It has been verified that lnc RNAs such as NONHSAT181139.1 and NONHSAT185097.1 are up-regulated in Hashimoto's thyroiditis cells;lnc RNAs such as NONHSAT152327.1,NONHSAT183385.1,NONHSAT213829.1 and NONHSAT245695.1 are down-regulated in Hashimoto's thyroiditis cells.3.Spearman analysis founds that lnc RNA: NONHSAT185097.1 was positively correlated with TPOAb,correlation coefficient was 0.469,P = 0.016(according to P =0.05 standard);lnc RNA: NONHSAT152327.1 is correlated with TPOAb and TGAb(P<0.05).