期刊论文

Daxx overexpression inhibits AngII-induced proliferation and migration in vascular smooth muscle cells

中文

南方医科大学学报

1673-4254

2019

39

10

1173-1179

10.12122/j.issn.1673-4254.2019.10.07

基金项目：国家自然科学基金（81673722，81603600）；湖南省教育厅重点 项目（16A156）；湖南省自然科学基金（2019JJ50426）；湖南中医药大学基 础医学一流学科开放基金（2018JCYX04）；校级研究生科研创新项目 （2018CX54） Supported by National Natural Science Foundation of China (81673722, 81603600). 作者简介：曹玉梅，硕士，E-mail: 861869487@qq.com 通信作者：庹勤慧，博士导师，教授，E-mail: qhtuo@aliyun.com

本校为第一机构

目的构建重组慢病毒表达载体p CDH-Daxx-EGFP,并探讨Daxx对血管紧张素Ⅱ（AngⅡ）诱导VSMCs增殖的影响。方法基于PAS（PCR-based Accurate Synthesis）的方法构建重组慢病毒表达载体pCDH-Daxx-EGFP。经测序和酶切验证后,用重组慢病毒表达载体pCDH-Daxx-EGFP与包装辅助载体共转染293T细胞进行慢病毒包装,收集病毒液纯化后感染VSMCs,Western blot法鉴定过表达Daxx的VSMCs株。然后将p CDH-EGFP病毒液感染的空载体细胞和pCDH-Daxx-EGFP病毒液的感染的过表达Daxx细胞都分成无血清培养基孵育组和AngⅡ孵育组。用MTT法检测细胞活性、流式细胞术观察细胞周期、划痕法检测细胞迁移、Western blot法检测细...

Objective To construct a recombinant lentiviral expression vector pCDH-Daxx-EGFP to investigate the effect of Daxx on the proliferation of vascular smooth muscle cells (VSMCs). Methods The recombinant lentiviral expression vector pCDHDaxx-EGFP was constructed using PCR-based accurate synthesis method. After identification by sequencing and enzyme digestion, the recombinant lentiviral vector was contransfected into 293T cells with lentivirus packaging vector. The recombinant lentivirus particles were collected and purified to infect VSMCs, whose...