| Objective: To investigate the expression of long non-coding RNA LIPE-AS1(LncRNA LIPE-AS1)in psoriasis vulgaris,and to explore effect of LIPE-AS1 on apoptosis and proliferation of Ha Ca T cells and the potential mechanism,to provide theoretical foundation for the precise treatment of psoriasis.Methods:(1)We used lncRNA array analysis to detect the differentially expressed lncRNAs in lesions and the adjacent skin of fifteen patients with psoriasis vulgaris.Based on lncRNA sequencing results,lncRNA LIPE-AS1,which is over expressed in lesions is selected.(2)Quantitative real-time reverse transcription-polymerase chain reaction(RT-PCR)was performed to validate the expression of lncRNA LIPE-AS1 in psoriasis lesions and adjacent skin in 5 patients with psoriasis vulgaris and normal skin in 5 healthy controls.(3)The downregulation of LIPE-AS1 expression was performed by the specific sh RNA lentiviral vectors.Lentiviral particles were infected into Ha Ca T cells,and cells with stable and down expression of LIPE-AS1 were selected by puromycin resistance.The expression of LIPE-AS1 was verified by RT-PCR.CCK-8 assay and flow cytometry were used to detect the effect of LIPE-AS1 on apoptosis and proliferation of Ha Ca T cells,and RT-PCR was used to detect the inflammatory cytokines in infected cells.(4)The possible target genes of LncRNA LIPE-AS1 was predicted by bioinformatics analysis,and the expression of the genes was detected by RT-PCR in 5 patients with PS and 5 healthy controls.And the expression of m RNA and protein of the genes in infected cells were detected by RT-PCR and Western blot.Results:(1)The results of lncRNA array analysis showed that,compared with adjacent skin,there were a large number of abnormally expressed lncRNAs in psoriatic lesions.(2)LncRNA LIPE-AS1 was significantly up-regulated in lesions of psoriasis vulgaris(P vlaue =1.277E-05,Fold Change =3.592).(3)Specific interference sh RNA-LIPE-AS1 lentivirus significantly inhibited LIPE-AS1 expression in Ha Ca T cells(p<0.05).(4)Knockdown of LIPE-AS1 induced G1/S cell cycle arrest,enhancesd apoptosis and promoted Ha Ca T cell proliferation.(5)In keratinocytes,knockdown of LIPE-AS1 inhibited INF-γ expression.(6)LncRNA LIPE-AS1 negatively regulated the expression of BCL2L2(BCL2 like 2).Conclusion: Compared with the adjacent skin and normal skin,LncRNA LIPE-AS1 is highly expressed in psoriastic lesions.Knockdown of LIPE-AS1 suppresses Ha Ca T cell proliferation by inducing G1/S cell cycle arrest and apoptosis,and the significant decrease of INF-γ expression.BCL2L2,a candidate target gene of LIPE-AS1,is upregulated in psoriasis lesions.And in the occurrence and development of psoriasis vulgaris,LIPE-AS1 overexpression may inhibits keratinocyte apoptosis and stimulates proliferation by inhibiting BCL2L2. |
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