Jiang Xiuli, Chen Jiajia, Song Quanjiang, Wang Weiling, Zhang Guangyan, Li Ye
Department of Neurology, People's Hospital of Rizhao, Rizhao, Shandong 276800, P.R. China.
Department of Clinical Laboratory, People's Hospital of Rizhao, Rizhao, Shandong 276800, P.R. China.
Exp Ther Med. 2017 Dec;14(6):6238-6242. doi: 10.3892/etm.2017.5345. Epub 2017 Oct 19.
The correlation between tuberous sclerosis complex 1 (TSC1) gene polymorphism and epilepsy was studied. In total, 38 patients with epilepsy treated in People's Hospital of Rizhao from May 2015 to June 2016 were selected as study subjects, as the observation group, 38 healthy people in the same period were selected as the control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to study the polymorphism of TSC1 gene in the above study subjects. The mRNA expression of TSC1 gene in the observation group and the control group was measured by fluorescence quantitative PCR, the expression of TSC1 protein in the control and observation group was measured by western blotting and ELISA. The polymorphisms of TSC1 gene in control group and observation group were analyzed by PCR-RFLP. There were three genotypes of TCS1 gene locus 142 in healthy population: CC (79.3%), CA (13.9%) and AA (6.8%), there were also three genotypes at locus 142 in the observation group: CC (21.3%), CA (26.4%) and AA (52.3%), there was significant difference in the genotypes at locus 142 between healthy population and the patients with epilepsy (P<0.05). It was observed by fluorescence quantitative PCR that there was no significant difference in the mRNA expression of TSC1 gene between the control group and the observation group (P>0.05). The expression of TSC1 gene was detected by western blot method. Western blotting showed no significant difference in TSC1 protein expression between the two groups (P>0.05). However, by determining the activity of TSC1 protein in the observation group and the control group by ELISA, it was found that TSC1 activity in healthy human body (8.95±2.41 U/ml) was much lower than that in the patients with epilepsy (29.27±4.06 U/ml), the difference was statistically significant (P<0.05). It was found that locus 142 may be located at the active center of TSC1 enzyme by homology modeling of SWISS-MODEL, the mutation of locus 142 could lead to the change of TSC1 activity. The polymorphism of locus 142 in TSC1 gene is correlated with epilepsy, that is, the increase of CA and AA content in locus 142 leads to the occurrence of epilepsy.
研究了结节性硬化症复合物1（TSC1）基因多态性与癫痫之间的相关性。选取2015年5月至2016年6月在日照市人民医院治疗的38例癫痫患者作为研究对象，即观察组，同期选取38例健康人作为对照组。采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法研究上述研究对象中TSC1基因的多态性。通过荧光定量PCR检测观察组和对照组中TSC1基因的mRNA表达，采用蛋白质免疫印迹法和酶联免疫吸附测定法检测对照组和观察组中TSC1蛋白的表达。通过PCR-RFLP分析对照组和观察组中TSC1基因的多态性。健康人群中TCS1基因位点142有三种基因型：CC（79.3%）、CA（13.9%）和AA（6.8%），观察组位点142也有三种基因型：CC（21.3%）、CA（26.4%）和AA（52.3%），健康人群与癫痫患者在位点142的基因型存在显著差异（P<0.05）。通过荧光定量PCR观察发现，对照组和观察组中TSC1基因的mRNA表达无显著差异（P>0.05）。采用蛋白质免疫印迹法检测TSC1基因的表达。蛋白质免疫印迹法显示两组间TSC1蛋白表达无显著差异（P>0.05）。然而，通过酶联免疫吸附测定法检测观察组和对照组中TSC1蛋白的活性，发现健康人体内TSC1活性（8.95±2.41 U/ml）远低于癫痫患者（29.27±4.06 U/ml），差异具有统计学意义（P<0.05）。通过SWISS-MODEL同源建模发现位点142可能位于TSC1酶的活性中心，位点142的突变可能导致TSC1活性的改变。TSC1基因位点142的多态性与癫痫相关，即位点142中CA和AA含量的增加导致癫痫的发生。
