基于3T3－L1细胞株的人类 ADRB1基因稳定过表达模型的构建

摘要

Objective:To establish an adipocyte model with stable overexpression of β1 -adrenergic receptor (ADRB1)gene.Methods:After the ligation of ADRB1 cDNA and lentiviral vector,the recombinant vector and pack-aging plasmids were co -transfected into 293T cells.Infection was performed with the medium.Stably transfected cells were selected with fluorescence -activated cell sorting(FACS).Results:There was a significant elevation of the level of ADRB1 protein in the overexpression group.Conclusion:The ADRB1 stable overexpression model has been estab-lished based on 3T3 -L1 cell line.%目的：构建 ADRB1基因稳定过表达的细胞模型。方法：将 ADRB1 cDNA 克隆片段和慢病毒载体连接；将重组载体与慢病毒包装质粒共转染293T 细胞，收集上清液感染3T3－L1前脂肪细胞，利用 GFP 流式分选法筛选稳转细胞株。结果：Western blot 检测证明，ADRB1过表达组的 ADRB1蛋白水平显著高于 mock 组。结论：基于3T3－L1细胞株的 ADRB1基因稳定过表达模型构建成功，为研究 ADRB1在肿瘤恶病质脂肪消耗中的作用奠定了基础。