Identification of CHO cell lines stably transfected by β1,β2 or β3 adrenoceptor gene respectively

ZENG Fan-xin; et al

doi:10.13406/j.cnki.cyxb.2011.03.041

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Identification of CHO cell lines stably transfected by β1,β2 or β3 adrenoceptor gene respectively

更新时间：2024-01-17

- Identification of CHO cell lines stably transfected by β1,β2 or β3 adrenoceptor gene respectively
- Journal of Chongqing Medical University Vol. 36, Issue 3, Pages: 323-326(2011)
- 作者机构：
  
  1. 重庆医药工业研究院创新药物研发部
  2. 重庆医科大学药学院药理教研室
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2011.03.041
  CLC： R965.1
- Published：2011，
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[1]曾凡新,董志,傅洁民,邓杰,刘晓海,杨佳丹.稳定表达β1、β2及β3肾上腺素受体细胞株的鉴定[J].重庆医科大学学报,2011,36(03):323-326.

ZENG Fan-xin, et al. Identification of CHO cell lines stably transfected by β1,β2 or β3 adrenoceptor gene respectively. [J]. Journal of Chongqing Medical University 36(3):323-326(2011)
[1]曾凡新,董志,傅洁民,邓杰,刘晓海,杨佳丹.稳定表达β1、β2及β3肾上腺素受体细胞株的鉴定[J].重庆医科大学学报,2011,36(03):323-326. DOI： 10.13406/j.cnki.cyxb.2011.03.041.

ZENG Fan-xin, et al. Identification of CHO cell lines stably transfected by β1,β2 or β3 adrenoceptor gene respectively. [J]. Journal of Chongqing Medical University 36(3):323-326(2011) DOI： 10.13406/j.cnki.cyxb.2011.03.041.

摘要

目的:对稳定转染有携带人β1、β2或β3肾上腺素受体基因质粒的CHO-K1细胞（β1-CHO、β2-CHO和β3-CHO细胞）进行鉴定

以为将这几种细胞用于β3肾上腺素受体激动剂的筛选奠定基础。方法:采用免疫细胞化学、免疫荧光、western blott和放射性配体结合法对3种细胞进行鉴定。结果:β1-CHO、β2-CHO和β3-CHO3种细胞经免疫细胞化学检测

细胞内均出现明显的棕黄色免疫阳性反应;经免疫荧光检测

3种细胞内均有明显的荧光;3种细胞提取的蛋白质经Western blot检测均出现预期的受体蛋白条带

且均可与不同浓度的放射性核素标记配体125I-cyanopindolol呈饱和结合。结论:β1-CHO、β2-CHO和β3-CHO细胞分别稳定表达有β1、β2或β3肾上腺素受体蛋白

可以用于特异性β3肾上腺素受体激动剂的筛选。

Abstract

Objective:To identify the CHO cell lines stably transfected by β1

β2 or β3 adrenergic receptor gene plasma respectively(named β1-CHO

β2-CHO or β3-CHO cell)

so that to use these cell lines for the screening of potential β3 agonists.Methods:The cell lines were identified by the methods of immunocytochemistry

immunofluorence

western blot and radioactive ligand binding assay.Results:β1-CHO

β2-CHO or β3-CHO cells showed positive staining by immunocytochemistry assay and obvious green fluorescence by immunofluoresence assay.The proteins extracted from these three cell lines showed predicted strap of β1

β2 or β3 receptor protein when detected by western blotting

and these cells can bind with a non-specific adrenergic radioligand 125I-cyanopindolol at a saturable binding way.Conclusion:β1-CHO

β2-CHO or β3-CHO cell lines stably expressed β1

β2 or β3 adrenoceptor separately and can be used for the screening of β3 adrenoceptor agonists.

关键词

肾上腺素受体β1肾上腺素受体β2肾上腺素受体β3肾上腺素受体激动剂

Keywords

adrenergic receptorβ1 adrenoceptorβ2 adrenoceptorβ3 adrenoceptoragonist

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