Differential Expression Of LncRNA TINCR In Psoriatic Lesion And Its Effect On Proliferation And Inflammation Of Keratinocyte

[Objective]1.With published data sets and bioinformatic methods,biological effect and character of differentially expressed genes in the tissues of psoriasis vulgaris were explored to have a better understanding of the pathogenesis of psoriasis.2.The expression level of the related differential LncRNA TINCR were verified by preliminary experiments using psoriatic lesions to provide relevant theoretical basis for further exploring its involvement in the pathogenesis and treatment of psoriasis.3.To explore the biological function of LncRNA TINCR in the process of proliferation of keratinocytes,which may provide a new therapeutic target for the treatment of psoriasis by interfering with LncRNA TINCR.4.To explore the effect of LncRNA TINCR on the production of inflammatory factors by keratinocytes.[Methods]1.Part Ⅰ:Bioinformatic methods were used to analyze and predict differentially expressed genes(DEGs).With integrated analysis of two data sets,differentially expressed LncRNAs in psoriatic lesions and differentially expressed LncRNAs involved in cultured keratinocyte differentiation respectively,differentially expressed genes(DEGs)related to psoriasis vulgaris were screened.LncRNA TINCR was selected as the main research object and purpose of this experimental study.2.Part Ⅱ:Study the expression level of the related differential LncRNA TINCR in the tissues of psoriasis vulgaris.Skin lesions of 10 patients with psoriasis vulgaris and skin tissues of 10 healthy controls were collected,which were divided into PV group(psoriasis skin tissue)and NN group(healthy human skin tissue).Real-time quantitative polymerase chain reaction(RT-qPCR)technology was used to detect the expression of LncRNA TINCR in each sample above,and the expression difference of LncRNA TINCR between two groups was estimated to further verify the results of bioinformatics analysis.3.Part Ⅲ:To investigate the biological function of LncRNA TINCR in the proliferation of keratinocytes.In human-immortalized keratinocyte HaCaT cells,knock down LncRNA TINCR by siRNA interference technology,and explore biological functions of LncRNA TINCR during HaCaT cell proliferation.MTT assay was used to assess HaCaT cell viability to study the effect of knockdown of LncRNA TINCR on the proliferation of HaCaT cells.4.Part Ⅳ:The effect of TINCR on the inflammation of keratinocytes:After transfecting Hacat cells with siRNA for 48h,ELISA was used to detect the serum levels of TNF-α,IL-1β and IL-6;Extract the total RNA and test the expression of TNF-α,IL-1β and IL-6 by RT-qPCR method.[Results]1.In part Ⅰ,117 genes with significant expression differences and transcriptome data sets with keratinocytes(KC)differentiation on days 0,3,and 6 were screened.By comparing and analyzing these two data sets,40 differentially expressed candidate genes related to psoriasis vulgaris were selected.29 were up-regulated and 11 were down-regulated.By comparing the expression levels of different LncRNAs in healthy skin tissue and psoriasis skin tissue,LncRNA TINCR was finally selected as a main research object2.In Pazrt Ⅱ,the results of RT-qPCR showed that,Compared with the control group,expression levels of LncRNA TINCR in the lesions of psoriasis vulgaris group was downregulated compared with the control group,with statistical differences(P<0.05),which verified the result of the bioinformatics analysis.3.In part Ⅲ,the results of MTT experiment showed that in siRNA-transfected HaCaT cells,the proliferation of Hacat cells transfected with si-TINCR was enhanced and promoted.4.The levels of TNF-α,IL-1β and IL-6 in the cell supernatant after transfection of Hacat cells with siRNA for 48h:the levels of IL-1β and IL-6 were significantly higher than those in the normal group(P<0.05).There is no significant difference in the level of TNF-α(P>0.05);Hacat cells knock down the total RNA of lncRNA TINCR samples,the expression of inflammatory factors TNF-α,IL-1β and IL-6 at the transcrition level:The mRNA expression of IL-1β and IL-6 was significantly higher than that of the negative control group(P<0.05).The mRNA expression of TNF-α had no significant difference compared with the control group(P>0.05).[Conclusions]1.Combined with bioinformatics analysis results,LncRNAs with significantly different differential expression related to psoriasis vulgaris screened.During which LncRNA TINCR was selected as the main research object in this paper.Its biological effects need to be further explored and studied.2.LncRNA TINCR was related to psoriasis and it may be involved in the development progression of psoriasis vulgaris by regulating keratinocytes,which is worthy of further study.3.It has been shown that LncRNA TINCR affected the pathogenesis of psoriasis vulgaris by inhibiting the process of proliferation of keratinocytes.4.LncRNA TINCR is involved in regulating the production of inflammatory factors IL-1β and IL-6 by keratinocytes.LncRNA TINCR may inhibit the inflammatory response of keratinocytes by inhibiting the release of related inflammatory factors.