Stable expression of the human TSH receptor in CHO cells and the functional assessment
10.3760/cma.j.issn.1000-6699.2018.02.012
- VernacularTitle:稳定表达全长人促甲状腺激素受体细胞株构建及功能评价
- Author:
Xiaofang TU
1
;
Hongmei ZHANG
;
Qing SU
Author Information
1. 200092,上海交通大学医学院附属新华医院内分泌科
- Keywords:
Human thyroid stimulating hormone receptor;
Cell line;
Establish;
Function
- From:
Chinese Journal of Endocrinology and Metabolism
2018;34(2):154-157
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish and assess a CHO (Chinese Hamster Ovary) cell line that stably expressed the human TSH receptor. Methods The gene of human TSH receptor was amplified by PCR from the recombinant plasmid that had been constructed previously,and then the target gene was linked to linearized vector GV208. Afterwards, with the reconstructed vector and helper vectors, the target gene was packaged into the lentivirus. The recombinant lentivirus infected the CHO cells and integrated the gene into the chromatin. The established CHO cell line was isolated and cultured and its function was assessed.Results The result of gene test identified the correct sequence of DNA of the transformant. The expression of mRNA in CHO cells transfected with the human TSHR was obviously higher than the cells in control or blank group. Within the range of certain density,the more bovine TSH was used to stimulate the established CHO,the more cAMP would be produced. When stimulated with serum of patients of Graves' disease, the cell line also significantly produced more cAMP. Conclusion The established CHO cell line transfected with the human TSHR can stably express the human TSH receptor and response to the stimulation of ligand of TSH receptor.
