miR-21通过PTEN/AKT减轻突变亨廷顿蛋白细胞毒性  

miR-21 alleviates the cytotoxicity of mutant huntingtin through PTEN/AKT

作　　者：李希凡 陈恬 方方 李和[2] LI Xi-fan;CHEN Tian;FANG Fang;LI He(Department of Human Anatomy,Guilin Medical University,Guilin 541004,Guangxi Province,China;Division of Histology and Embryology,Department of Anatomy,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,Hubei Province,China)

机构地区：[1]桂林医学院人体解剖学教研室,广西桂林541004 [2]华中科技大学同济医学院人体解剖学系组织胚胎学教研室,武汉430030

出　　处：《中国临床解剖学杂志》2020年第1期51-56,共6页Chinese Journal of Clinical Anatomy

基　　金：广西自治区自然科学基金青年项目（2017GXNSFB A198155）;广西科技基地和人才专项（桂科AD18281011）;广西脑与认知神经科学重点实验室开放课题（GKLBCN-20180105-06）

摘　　要：目的探讨miR-21减轻突变亨廷顿蛋白(mutant huntingtin,mHtt)细胞毒性的机制。方法利用qRT-PCR检测miR-21在亨廷顿病(Huntington’s disease,HD)转基因小鼠脑组织以及HEK293-160Q细胞中的变化;双荧光素酶实验检测PTEN是否为miR-21的靶基因;转染miR-21 mimics后,CCK8检测细胞活力,Caspase-3活性酶标检测Caspase-3活性,Western blotting检测PTEN、Ser-473位点磷酸化的AKT以及AKT。结果qRT-PCR结果显示,与野生型小鼠及HEK293-20Q细胞相比,HD转基因小鼠脑组织及HEK293-160Q细胞中miR-21均显著降低;双荧光素酶实验证实PTEN为miR-21靶基因;在HEK293-160Q细胞中转染miR-21 mimics后PTEN显著降低、p-AKT-Ser 473/AKT显著升高。结论miR-21能通过降低PTEN来提高p-AKT-Ser 473/AKT,减轻mHtt所引起的细胞毒性继而提高细胞活力、抑制细胞凋亡。Objective To detect the mechanism of miRNA-21 in reducing cytotoxicity of mutant Huntingtin(mHtt).Methods The changes of miRNA-21 in brain tissues of Huntington’s disease(HD)transgenic mice and HEK293-160 Q cells were detected by quantitative real-time polymerase chain reaction(qRT-PCR);Whether PTEN was the target gene of miRNA-21 or not was identified by double luciferase test;after transfected the miRNA-21 mimics,cell viability was detected by CCK8,caspase-3 activity was determined by caspase-3 activity assay kit;PTEN,phosphorylated-Ser-473 AKT and AKT were detected by Western Blotting.Results qRT-PCR result showed that the levels of miRNA-21 in brain tissues of HD transgenic mice and HEK293-160 Q cells were significantly decreased compared with wild-type mice and HEK293-20 Q cells;PTEN was the target gene of miRNA-21 which was confirmed by double luciferase test;Western Blotting showed that PTEN decreased significantly and p-AKT-Ser 473 AKT/AKT significantly increased in HEK293-160 Q cells after transfected the miRNA-21 mimics.Conclusions miR-21 can increase the p-AKT-Ser 473/AKT by decreasing the PTEN,increase the cell viability and inhibit the apoptosis by alleviating the cytotoxicity of mHtt.

关 键 词：突变亨廷顿蛋白 MIR-21 PTEN AKT 

分 类 号：R741[医药卫生—神经病学与精神病学]