| Objective.To investigate the association between the GTF2 I rs117026326 polymorphisms and patients with multiple sclerosis(MS)and neuromyelitis optica spectrum disorder(NMOSD).Subjects and Methods.We conducted a case-control study which included 96 patients with NMOSD,106 patients with MS and 930 healthy control group.All subjects were patients who admitted to the Department of Neurology and the Neuroscience Center at The First Hospital of Jilin University between 2015 and 2018.Patients with MS were diagnosed by neurologists according to the revised Mc Donald criteria(Polman et al.,2011).Patients with NMOSD met the revised diagnostic criteria for NMOSD(Wingerchuk et al.,2015).A total of 930 healthy controls were recruited from the staff of the same hospital to be genotyped and compared with the patients as part of a case-control study.All patients and healthy control subjects provided written informed consent.1.Genomic DNA samples were extracted using a blood/tissue DNA magnetic bead extraction kit.For the selected SNPs,designed Taq Man SNP genotyping assays were used.The sequences of primers and probes synthesized follows: rs117026326,forward primer: CTG TTT TCT GTT TTA GGT TAG TTT GCA;reverse primer: CAA CGC TGT GGA TGA ATT TCA;risk probe: FAMACT ATT TTC ATG GGC TGG-MGB;non-risk probe: HEX-ACT ATT TTC ATG GGC CGGMGB.2.Peripheral blood mononuclear cells(PBMCs)were isolated from peripheral blood from patients with MS and NMOSD.Total RNA was extracted with Trizol according to the manufacturer’s instructions.RNA reverse transcription c DNA and QPCR were used to determine the expression level of GTF2 I and its nearby gene GTF2IRD1 m RNA.The m RNA expression of GTF2 I and GTF2IRD1 in each sample was determined using real-time quantitative reverse transcription PCR and was normalized to the GAPDH control.3.Statistical analysis was then performed using Plink,version 1.90.Results.We observed a significant genetic association between the variant rs117026326 and NMOSD(P = 3.00×10-8,OR = 2.534),but the association with MS was not significant(P = 0.651,OR = 1.088).Gene expression analyses showed that there was no significant association between the messenger RNA(m RNA)expression of GTF2 I,GTF2IRDI and genotypes at the variant.Conclusion.The results show that the GTF2 I rs117026326 T allele increase risk of NMOSD,but not with MS.There are no significant associations between the genotypes of the variant rs117026326 and GTF2 I,GTF2IRD1 expression.Our findings shed new light on the pathogenesis of NMOSD independent of MS,and suggest that the two diseases may have different genetic risk factors. |
