Study On The Role Of Long Non-coding RNA In Peripheral Blood T Cells Of Systemic Lupus Erythematosus

ObjectiveThe expressions of 7 long non-coding RNA(lnc RNA)(LINC02446,SNHG16,PRKCQ-AS1,RFX3-AS1,MIR22 HG,MIF-AS1,SNHG17)in T cells of systemic lupus erythematosus(SLE)patients and normal controls were analyzed in this study,and the correlations between the levels of lnc RNA and the clinical manifestations,laboratory indexes and drug therapy of SLE patients were discussed.The values of lnc RNA as diagnostic biomarkers for SLE disease were explored.Finally,lnc RNA,which was differentially expressed and associated with clinical indicators and can be used as a diagnostic marker for SLE disease,was selected for follow-up cells function experiment studies to explore the influence of its over-expression on T cells function.MethodsThe case-control study was designed in this study,involving a total of 99 SLE patients and 118 healthy controls.After obtaining informed consent,collected 10 ml peripheral venous blood of the study subjects to obtain peripheral blood mononuclear cells(PBMCs),and T cells were got from PBMCs.After extraction of total RNA from T cells,complementary DNA(c DNA)was reversely transcribed from total RNA.The quantitative reverse transcription polymerase chain reaction(q RT-PCR)assay was used to determine the relative expression levels of candidate lnc RNA in the SLE group and control group,and to compare the differences in expressions.t test or Mann-Whitney U test were used to analyze the associations between lnc RNA expression levels and clinical indicators according to data types.The values of differentially expressed lnc RNA as diagnostic markers of SLE were determined by receiver operating characteristic(ROC)curve.LINC02446 was stably over-expressed in Jurkat(clone E6-1)cells by constructing lentiviral vector.Annexin V-APC/7-AAD double staining and CCK-8 method were used to observe the effect of LINC02446 over-expression on T cells proliferation and apoptosis.ResultsCompared with healthy controls,LINC02446 and MIR22 HG levels were upregulated(both P<0.05),while PRKCQ-AS1,SNHG16,RFX3-AS1,MIF-AS1 and SNHG17 levels were downregulated(all P<0.05)in T cells of SLE patients.The expressions of LINC02446,SNHG16 and SNHG17 in SLE patients with lupus nephritis(LN)were higher than those patients without LN(Z=-3.753,P<0.001;Z=-2.718,P=0.007;Z=-2.662,P=0.008);The expression level of MIF-AS1 was higher in SLE patients with fever(Z=-2.006,P=0.045).There were statistical differences among SLE patients with systemic lupus erythematosus disease activity index(SLEDAI)≤4 group and SLE patients with SLEDAI>4 group.RFX3-AS1 and MIR22 HG expressions were higher in SLE patients with SLEDAI>4 group(all P<0.05);LINC02446 and MIR22 HG levels were higher in the anemia group(Z=-2.186,P=0.029;Z=-2.882,P=0.004);In the group with erythropenia,LINC02446(Z=-2.435,P=0.015),SNHG16(t=-2.382,P=0.019),MIR22HG(Z=-3.013,P=0.003)and SNHG17(Z=-2.312,P=0.021)were highly expressed.The expression level of LINC02446(Z=-2.227,P=0.026),SNHG16(Z=-2.715,P=0.007)and SNHG17(Z=-2.712,P=0.007)in patients with proteinuria were higher than those in patients without proteinuria.The AUC values of PRKCQ-AS1 and LINC02446 as biomarkers of SLE were both above 0.7,indicating good diagnostic value.When ROC analysis was performed in pairwise combinations,the results showed that the diagnostic value of PRKCQ-AS1 combined with LINC02446 was the highest,the AUC(95% CI)was 0.8615(0.8117-0.9113),the sensitivity was 83.90% and the specificity was 76.77%.The AUC value of LINC02446 and its combination with SNHG16 in determining whether SLE patients suffer from LN were above 0.7,showing good diagnostic value,among which LINC02446 combined with SNHG16 had the highest diagnostic value for LN.The AUC(95 CI)value was 0.7244(0.6216-0.8272),the sensitivity was 81.82% and the specificity was 65.45%.Further cells functional studies showed that over-expression of LINC02446 led to increases in the percentages of T cells apoptosis(both early and late apoptotic stage)(P<0.05).At the same time,the proliferative ability of T cells(24 h,48 h,72 h)was decreased(P<0.05).ConclusionsLINC02446,SNHG16,PRKCQ-AS1,RFX3-AS1,MIR22 HG,MIF-AS1 and SNHG17 were abnormally expressed in T cells of SLE patients,and the expression levels of lnc RNA were associated with clinical features.LINC02446 can be used to distinguish SLE patients from healthy individuals.LINC02446 may be involved in the progression of SLE by influencing T cells function.