Construction and drug resistance evaluation of cell strains which stably express FIP1L1-PDGFRA protein and its mutants

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 evaluate 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 sensitivity 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.

Keywords

• fip1l1-pdgfra fusion gene|gene mutation|lentiviral vector|baf3 cell|chronic eosinophilic leukemia