Association Study Of TGF-β1 Gene Polymorphisms And Its MRNA Level With LncRNA In Patients With Systemic Sclerosis

Objective:To explore the role of transforming growth factor-β1(TGF-β1)gene polymorphisms in systemic sclerosis(SSc)risk among the Han people in Anhui,China.Methods:A hospital-based case-control study was used,including SSc patients(n=100)and healthy volunteers(n=100).TGF-β1 gene were genotyped using improved multiplex ligase detection reaction(i MLDR)method.Before and after adjusting for confounding factors,the genotypes and allele frequencies of the both groups were calculated respectively,and the difference of dominant and recessive models between SSc and control individuals was studied.Additionally,haplotypes were constructed after linkage disequilibrium(LD)analysis.Results:Two SNPs of TGF-β1(rs1800469 and rs1800470)were studied.The genotype frequencies in both groups were satisfied by the Hardy-Weinberg Equilibrium(HEW).No matter whether the potential confounding factors(age and gender)were controlled or not,logistic regression analysis suggested that the genotype and allele frequencies of rs1800469 and rs1800470 had no statistical difference between SSc cases and healthy individuals(all P>0.05).Through genetic model analysis,no significant difference between the dominant model(GG+GA vs AA)(before:OR=1.247,95%CI=0.650-2.392;after:OR=1.271,95%CI=0.659-2.450)and the recessive model(GG vs GA+AA)(before:OR=1.000,95%CI=0.517-1.934;after:OR=1.005,95%CI=0.517-1.954)of rs1800469 before and after adjusting for potential confounders(e.g.age and gender).Also,we did not find any significant association of rs1800470 with SSc risk(all P>0.05).χ~2test found that the three genotypes and alleles frequencies of rs1800469 and rs1800470 were not related to the clinical or laboratory manifestations of SSc patients,such as skin stiffness,joint involvement,organ involvement,C3 and antinuclear antibody(ANA)(all P>0.05).LD analysis was conducted,there was high LD between the chosen SNPs rs1800469 and rs1800470(D’=0.99,r~2=0.98).Furthermore,haplotype analysis was used through the distribution of haplotypes in SSc and healthy controls,these two haplotypes frequencies(AG and GA)were not significantly different in two group(all P>0.05).Conclusion:This study preliminary revealed polymorphism of TGF-β1 gene(rs1800469 and rs1800470)was not statistical significant with SSc risk in Chinese Han population in Anhui,nor was it correlated with clinical manifestations and laboratory characteristics of SSc patients.Objective: To identify the relationship of long non-coding RNA(lnc RNA)and transforming growth factor-β1(TGF-β1)mRNA in peripheral blood mononuclear cells(PBMC)of systemic sclerosis(SSc)patients.Methods: We performed a case-control study of 41 SSc cases and 42 healthy individuals.Participants’ peripheral blood(5ml)was extracted.The expression levels of molecules were measured by q RT-PCR.The relationship between lnc RNAs and TGF-β1mRNA was performed by Spearman rank correlation.Results: No significant difference were detected in age and gender between the SSc and healthy individuals.The results found that the expressions of H19,MALAT1 and TGF-β1 mRNA in PBMC of SSc patients were 1.881(0.801,4.419),1.290(0.427,4.619)and 0.785(0.481,1.671),respectively by Mann-Whitney U nonparametric test.However,the expression level of H19 was raised significantly compared to HC(P=0.008).No significant different between SSc patients and normal controls regarding the expression of MALAT1 and TGF-β1 mRNA(P=0.334,P=0.536,respectively).Furthermore,the correlation between the expression levels of H19,MALAT1 and clinic indicators among SSc patients were analyzed.The expression level of H19 in the C-reactive protein(CRP)elevated group was lower than that in reference group(P=0.047).In addition,a positive association was revealed between the expression level of lnc RNAs and TGF-β1 mRNA(rs=0.750 P<0.001;rs=0.381 P=0.014,respectively).Conclusion: The expression level of H19 in PBMC of SSc cases was elevated than that of healthy volunteer,and was associated with CRP,which indicated H19 may be related to inflammation in the early stage of SSc illness,whereas MALAT1 may not be directly involved in the risk of SSc.Among SSc cases,there was a positive correlation between lnc RNA and TGF-β1 mRNA expression levels,suggesting that MALAT1,H19 may be involved in the pathogenesis of SSC by affecting TGF-β1 levels.