Construction of a Differentiated Embryo Chondrocyte 1 Lentiviral Expression Vector and Establishment of its Stably Transfected HGC27 Cell Line

Abstract

AbstractHuman differentiated embryo chondrocyte 1 (DEC1), has been suggested to play key roles in immune regulation, cell differentiation, proliferation and apoptosis, circadian rhythms, hypoxia response and carcinogenesis. However, the role of DEC1 in gastric cancer have not been well established. Lentiviral vectors are widely used for the stable expression of genes and currently under development for clinical use in gene therapy. Therefore we intended to construct a lentiviral DEC1 expression vector and then establish a gastric cancer cell line with stable expression of DEC1. The coding sequence of gene was amplified using PCR and cloned into pGV218 vector. 293T cells were transfected using Lipofectamine 2000 and packaged for the recombinant lentivirus particles. When the cloned sequence was identified to be right, the recombinant lentivirus particles were amplified in a large quantity. The titer of virus was determined by real-time PCR. Subsequently, we collected the lentivirus venom to infect the HGC27 cells and establish a stable, overexpressed cell line named GFP/DEC1-HGC27. This study will provide a new cell model for further study of the role of DEC 1in the pathogenesis of gastric cancer.KeywordsDEC1Lentiviral expression vectorGastric cancerHGC27