Construction and drug resistance evaluation of cell strains which stably express FIP1L1-PDGFRA protein and its mutants
10.16352/j.issn.1001-6325.2024.01.0063
- VernacularTitle:稳定表达FIP1L1-PDGFRA及其突变体细胞株的构建和耐药性评价
- Author:
Jingwen LIU
1
;
Haoxin SONG
;
Lei ZHU
Author Information
1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 中国医学科学院医学表观遗传研究中心 药理学系,北京 100005
- Keywords:
FIP1L1-PDGFRA fusion gene;
gene mutation;
lentiviral vector;
BaF3 cell;
chronic eosinophilic leukemia
- From:
Basic & Clinical Medicine
2024;44(1):63-68
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct mouse BaF3-FIP1L1-PDGFRA(F/P),BaF3-F/P-T674I and BAF3-F/P-D842V pre-B cell strains which stably express F/P fusion protein and its T674I and D842V mutants in order to e-valuate their activity by checking their responses to tyrosine kinase inhibitors(TKIs).Methods Lentivirus infected technique was used to transfect the target gene into BaF3 cells.RT-qPCR was used to detect mRNA expression,and CCK-8 method was used to detect the inhibitory effect of TKIs on the proliferation of stable cell strains.Results The constructed BaF3-F/P,BaF3-F/P-T674I and BAF3-F/P-D842V cell strains all transcripted FIP1L1 and PDGFRA mRNA.They exhibited malignant phenotypic characteristics of proliferation independent of IL-3 and sen-sitivity to corresponding TKIs.Conclusions The pre-B-cell strains stably expressing F/P and its T674I and D842V mutants are successfully constructed,which provide a good cell model for the development of compounds targeting at those molecules.
