miR-21通过PTEN/AKT减轻突变亨廷顿蛋白细胞毒性

doi:10.13418/j.issn.1001-165x.2020.01.011

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (1): 51-56.doi: 10.13418/j.issn.1001-165x.2020.01.011

miR-21通过PTEN/AKT减轻突变亨廷顿蛋白细胞毒性

李希凡¹，　陈恬¹，　方方¹，　李和²

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

收稿日期:2019-04-23 出版日期:2020-01-25 发布日期:2020-02-18
通讯作者: 方方，副教授，研究生导师，E-mail：fangdouble@163.com；李和，教授，E-mail：heli_tjmu@163.com
作者简介:李希凡（1996-），男，在读硕士，研究方向：神经疾病的细胞与分子生物学研究，E-mail: 603418409@qq.com
基金资助:
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）

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

LI Xi-fan¹, CHEN Tian¹, FANG Fang¹, LI He²

1. Department of Human Anatomy, Guilin Medical University, Guilin 541004, Guangxi Province, China; 2.Division of Histology and Embryology, Department of Anatomy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030,Hubei Province, China

Received:2019-04-23 Online:2020-01-25 Published:2020-02-18

摘要/Abstract

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。

关键词: 突变亨廷顿蛋白, 　miR-21, 　PTEN, 　AKT

Abstract: 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 HEK293-160Q 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 HEK293-160Q cells were significantly decreased compared with wild-type mice and HEK293-20Q 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 HEK293-160Q 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.

Key words: Mutant huntingtin, 　miR-21, 　PTEN, 　AKT

中图分类号:

R741

李希凡, 陈恬, 方方, 李和. miR-21通过PTEN/AKT减轻突变亨廷顿蛋白细胞毒性[J]. 中国临床解剖学杂志, 2020, 38(1): 51-56.

LI Xi-fan, CHEN Tian, FANG Fang, LI He. miR-21 alleviates the cytotoxicity of mutant huntingtin through PTEN/AKT[J]. Chinese Journal of Clinical Anatomy, 2020, 38(1): 51-56.

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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

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