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Abstract：
The drug-resistant temporal lobe epilepsy (TLE) has recently been associated with single nucleotide variants (SNVs) in microRNA(miR)-146a (MIR-146A) (rs2910164) and Sodium Voltage-Gated Channel Alpha Subunit 1 (SCN1A) (rs2298771 and rs3812718) genes. Moreover, no studies have shown an association between these SNVs and susceptibility to drug-resistant and drug-responsive TLE in Brazil. Thus, deoxyribonucleic acid (DNA) samples from 120 patients with TLE (55 drug-responsive and 65 drug-resistant) were evaluated by real-time polymerase chain reaction (RT-PCR). A total of 1171 healthy blood donor individuals from the Online Archive of Brazilian Mutations (ABraOM, from Portuguese Arquivo Brasileiro On-line de Mutações), a repository containing genomic variants of the Brazilian population, were added as a control population for the studied SNVs. MIR-146A and SCN1A relative expression was performed by quantitative RT-PCR (qRT-PCR). The statistical analysis protocol was performed using an alpha error of 0.05. TLE patient samples and ABraOM control samples were in Hardy-Weinberg equilibrium for all studied SNVs. For rs2910164, the frequencies of the homozygous genotype (CC) (15.00% vs. 9.65%) and C allele (37.80% vs. 29.97%) were superior in patients with TLE compared to controls with a higher risk for TLE disease [odds ratio (OR) = 1.89 (95% confidence interval (95%CI) = 1.06-3.37); OR = 1.38 (95%CI = 1.04-1.82), respectively]. Drug-responsive patients also presented higher frequencies of the CC genotype [21.81% vs. 9.65%; OR = 2.58 (95%CI = 1.25-5.30)] and C allele [39.09% vs. 29.97%; OR = 1.50 (95%CI = 1.01-2.22)] compared to controls. For rs2298771, the frequency of the heterozygous genotype (AG) (51.67% vs. 40.40%) was superior in patients with TLE compared to controls with a higher risk for TLE disease [OR = 2.42 (95%CI = 1.08-5.41)]. Drug-resistant patients presented a higher AG frequency [56.92% vs. 40.40%; OR = 3.36 (95%CI = 1.04-17.30)] compared to the control group. For rs3812718, the prevalence of genotypes and alleles were similar in both studied groups. The MIR-146A relative expression level was lower in drug-resistant compared to drug-responsive patients for GC (1.6 vs. 0.1, p-value = 0.049) and CC (1.8 vs. 0.6, p-value = 0.039). Also, the SCN1A relative expression levels in samples from TLE patients were significantly higher in AG [2.09 vs. 1.10, p-value = 0.038] and GG (3.19 vs. 1.10, p-value < 0.001) compared to the AA genotype. In conclusion, the rs2910164-CC and rs2298771-AG genotypes are exerting significant risk influence, respectively, on responsive disease and resistant disease, probably due to an upregulated nuclear factor kappa B (NF-kB) and SCN1A loss of function.
摘要：
最近，耐药性颞叶癫痫（TLE）与microRNA（miR）-146a（MIR-146A）（rs2910164）和钠电压门控通道α亚基1（SCN1A）（rs2298771和rs3812718）基因中的单核苷酸变体（SNV）相关。此外，在巴西,没有研究表明这些SNV与耐药和药物反应性TLE的易感性之间存在关联.因此，通过实时聚合酶链反应（RT-PCR）评估了120例TLE患者（55例药物反应性和65例耐药）的脱氧核糖核酸（DNA）样本。来自巴西突变在线档案（ABraOM，来自葡萄牙ArquivoBrasileiroOn-linedeMutaçšes)，一个包含巴西人口基因组变异的储存库，被添加为研究的SNV的对照种群。通过定量RT-PCR(qRT-PCR)进行MIR-146A和SCN1A相对表达。使用0.05的α误差进行统计分析方案。对于所有研究的SNV,TLE患者样品和ABraOM对照样品处于Hardy-Weinberg平衡。对于rs2910164，纯合基因型(CC)的频率(15.00%vs.9.65%）和C等位基因（37.80%vs.29.97％）在TLE患者中优于TLE疾病风险较高的对照组[比值比（OR）=1.89（95％置信区间（95CI）=1.06-3.37）；OR=1.38（95CI=1.04-1.82），分别]。药物反应性患者的CC基因型频率也较高[21.81％vs.9.65%；OR=2.58（95CI=1.25-5.30）]和C等位基因[39.09%vs.29.97%；与对照组相比，OR=1.50(95CI=1.01-2.22)]。对于rs2298771，杂合基因型(AG)的频率(51.67%vs.40.40％）在TLE患者中优于TLE疾病风险较高的对照组[OR=2.42（95CI=1.08-5.41）]。耐药患者的AG频率较高[56.92%vs.40.40%；与对照组相比，OR=3.36(95CI=1.04-17.30)]。对于rs3812718，两个研究组的基因型和等位基因患病率相似。与GC的药物反应性患者相比，耐药患者的MIR-146A相对表达水平较低（1.6vs.0.1，p值=0.049）和CC（1.8vs.0.6，p值=0.039）。此外，来自TLE患者的样本中的SCN1A相对表达水平在AG中明显更高[2.09vs.1.10，p值=0.038]和GG（3.19vs.1.10，p值<0.001）与AA基因型相比。总之,rs2910164-CC和rs2298771-AG基因型具有显著的风险影响，分别,关于反应性疾病和抗性疾病，可能是由于核因子κB（NF-kB）和SCN1A功能的上调。
