Abstract
Objective To construct an Esophageal cancer Eca109 cell line which can stably overexpressing the ubiquitin specific peptidase 53 (USP53). Methods The USP53 full-length gene was amplified by Polymerase Chain Reaction (PCR) using a plasmid containing the full-length sequence of USP53 purchased by GE. The constructed USP53 lentiviral plasmid and packaging plasmid were co-transfected into 293T cells to obtain concentrated lentiviral particles, and then the lentiviral particles were infected with Eca109 cells. The medium containing puromycin was selected for stable transfected cell lines. Results The mRNA expression of USP53 in overexpressing cell lines (45.070±0.942) was significantly higher than that in the control group (1.091±0.075) and the normal group (1.000±0.023). The difference was statistically significant (P<0.05), Western blotting showed that the expression of the tagged protein HA was significant expressed in the overexpressed cell line, and no tagged protein expression was observed in the control group and the normal group. Conclusion Human ubiquitin specific peptidase 53 overexpressing cell line was successfully constructed. Key words: Esophageal cancer; Eca109; Ubiquitin specific peptidase 53; Lentivirus
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