Construction of recombinant antisense Smad3 adenoviral vector and its expression in vitro

Objective To construct the recombinant antisense Smad3 adenoviral vector with efficient expression in cuhured cells in vitro, and to evaluate the possibility of gene therapy for pathologic scar. Methods The recombinant antisense Smad3 adenoviral vector was constructed by the direct DNA cloning protocol and then transfected into 293 cells for virus packaging. After amplification and purification, the recombinant adenovirus was used to infect the keloid fibroblasts. The antisense Smad3 mRNA transcription of the infected cells was detected by RT-PCR. Results The recombinant adeno-antiSmad3 was correctly constructed and confirmed by both restriction analysis and PCR analysis. RT-PCR showed the expression of adenovirus mediated antiSmad3 mRNA in keloid cells. Conclusion These results demonstrate that the recombinant antisense Smad3 adenoviral vector can be expressed in cuhured keloid fibroblasts in vitro, and it may provide a new therapeutic strategy for keloid gene therapy.