Construction of adenovirus vector harboring RANTES regulated by oxygen-dependent degradation domain and its chemoattractant activity in vitro / 第二军医大学学报

ABSTRACT

Objective: To construct an adenovirus vector harboring the human RANTES gene regulated by oxygen-dependent degradation domain (ODD) and to observe its chemoattractant activity in vitro. Methods: The human RANTES gene was fused with ODD by PCR and the recombinant adenovirus was used to construct SG511-CCL5-ODD with the Gateway System. Viral replication experiments were performed to evaluate the selective replication ability of SG511-CCL5-ODD. The expression of RANTES protein was determined by ELISA under normal and hypoxia condition. Chemotactic test was used to analyze the chemoattractant ability of the expressed RANTES in liver cancer cells. Results: A recombinant adenovirus SG511-CCL5-ODD was constructed successfully. Cells infected with the recombinant virus expressed RANTES selectively. The expression of RANTES protein in the transfected liver cancer cells was higher under hypoxia condition than under normal condition (P<0.05), indicating ODD can effectively regulate RANTES protein expression. Chemotactic test showed that liver cancer cell infected with SG511-CCL5-ODD had the ability to recruit NK92 cells. Conclusion: Recombinant vector SG511-CCL5-ODD can effectively infect liver cancer cell line HepG2 and Hep3B, and can express RANTES protein under the regulation of ODD, demonstrating a chemoattractant activity in vitro.