Laboratory of Cell and Molecular Tumor Biology and Bioactive Compounds, São Francisco University Medical School (USF), Bragança Paulista, São Paulo, Brazil.
Department of Pathology, University of Campinas (Unicamp), Campinas, São Paulo, Brazil.
Epilepsy Res. 2020 May;162:106305. doi: 10.1016/j.eplepsyres.2020.106305. Epub 2020 Feb 29.
The aim of this study was to evaluate single nucleotide variants (SNVs) n.-411A > G (rs57095329) and n.60 G > C (rs2910164) in microRNA (miR)-146a, related to suppressing of TRAF6 with risk for epilepsy, as well as miR-146a and TRAF6 levels.
DNAs were extracted from epileptogenic tissues and blood leukocytes from drug-resistant epilepsy patients and healthy-individuals, respectively. Genotypes were identified by real-time PCR. Hardy-Weinberg equilibrium (HWE) and Fisher or X tests evaluated the difference between groups. The disease risk was assessed by odds ratio (OR) with 95 % confidence interval (95 %CI). The prognostic impact on probability seizure-free survival (PSF) was evaluated by Kaplan-Meier and log-rank tests.
For rs57095329 both control and patient samples were not in HWE (p < 0.05) and the genotypes prevalence was similar in patients and controls (p>0.05). For rs2910164, control samples were in HWE (p = 0.61), contrasting with patients (p = 0.03), and similar frequencies of wild-type homozygous (GG) (43.4 % vs. 34.4 %, p = 0.2) and variant (CC) genotypes (8.0 % vs. 6.6 %, p = 0.6) were observed in patients and controls, respectively. However, increased frequency of heterozygous (GC) was observed in patients compared to controls (59.0 % vs. 42.7 %, p = 0.04) with 1.98 (95 %CI=1.09-3.57) risk for epilepsy. The miR-146a expression level in the epileptogenic tissues was lower in the GC (p = 0.02) and CC (p = 0.09) compared to GG genotype. TRAF6 expression level was higher in CC than in GG genotype (p = 0.09). Interestingly, there was an increased frequency of patients harboring GC genotype and less time until surgery compared to patients harboring GG or CC (36.06 % vs. 11.5 %, p = 0.01), confirmed by PSF (p = 0.04).
The GC genotype for SNV rs2910164 appears associated with susceptibility to drug-resistant epilepsy due to the decreased MIR146a expression, favoring NF-kB pathway through TRAF6.
本研究旨在评估微 RNA（miR）-146a 中与 TRAF6 抑制相关的单核苷酸变异（SNV）n.-411A > G（rs57095329）和 n.60G > C（rs2910164），以及 miR-146a 和 TRAF6 水平与癫痫风险的关系。
从耐药性癫痫患者的致痫组织和白细胞中提取 DNA，分别采用实时 PCR 法鉴定基因型。采用 Hardy-Weinberg 平衡（HWE）和 Fisher 或 X 检验评估组间差异。采用比值比（OR）和 95 %置信区间（95 %CI）评估疾病风险。采用 Kaplan-Meier 和对数秩检验评估对无癫痫发作生存概率（PSF）的预后影响。
rs57095329 时，对照组和患者样本均未达到 HWE（p < 0.05），且患者和对照组的基因型患病率相似（p > 0.05）。rs2910164 时，对照组符合 HWE（p = 0.61），而患者组不符合（p = 0.03），患者和对照组野生型纯合子（GG）（43.4 %比 34.4 %，p = 0.2）和变异型（CC）基因型（8.0 %比 6.6 %，p = 0.6）的频率相似。然而，与对照组相比，患者中杂合子（GC）的频率增加（59.0 %比 42.7 %，p = 0.04），癫痫的风险为 1.98（95 %CI=1.09-3.57）。致痫组织中 miR-146a 的表达水平在 GC（p = 0.02）和 CC（p = 0.09）基因型中低于 GG 基因型。CC 型的 TRAF6 表达水平高于 GG 型（p = 0.09）。有趣的是，与 GG 或 CC 基因型相比，携带 GC 基因型的患者的手术时间较短（36.06 %比 11.5 %，p = 0.01），无癫痫发作生存概率（p = 0.04）也得到了证实。
SNV rs2910164 的 GC 基因型与耐药性癫痫的易感性有关，原因是 miR146a 表达降低，通过 TRAF6 促进 NF-kB 通路。
