The transcription factor nuclear factor kB (NF-kB) plays a key role in the delayed xenograft rejection (DXR). One of the important objects in the field is how to inhibit the NF-kB activity at optimal level. Thus, a mo...The transcription factor nuclear factor kB (NF-kB) plays a key role in the delayed xenograft rejection (DXR). One of the important objects in the field is how to inhibit the NF-kB activity at optimal level. Thus, a modified E1A gene (E1AD) containing function domain (1—80 aa) and nuclear localization domain (139—243 aa) was used and cloned into an eucaryotic expression vector pcDNA3 to transfect the porcine aortic endothelial cells (PAEC). The stable transfectants were screened with G418. E1AD gene was able to be stably expressed in the PAEC and could not affect the growth of PAEC as analyzed by RT-PCR and cell growth rate. Reporter gene assay demonstrated that E1AD was capable of inhibiting NF-kB activity in the PAEC in-duced by TNF-a without sensitizing to apoptosis, and the rate of inhibition was 53%. Furthermore, E1AD inhibited the expression of a NF-kBdependent inflammatory gene E-selectin in the cells, and the rate of inhibition was 63%. In summary, the usage of E1AD gene may be a new strategy to overcome DXR in the xenotransplantation.展开更多
基金This work was supported by Beijing "248"High-Tech Program China.
文摘The transcription factor nuclear factor kB (NF-kB) plays a key role in the delayed xenograft rejection (DXR). One of the important objects in the field is how to inhibit the NF-kB activity at optimal level. Thus, a modified E1A gene (E1AD) containing function domain (1—80 aa) and nuclear localization domain (139—243 aa) was used and cloned into an eucaryotic expression vector pcDNA3 to transfect the porcine aortic endothelial cells (PAEC). The stable transfectants were screened with G418. E1AD gene was able to be stably expressed in the PAEC and could not affect the growth of PAEC as analyzed by RT-PCR and cell growth rate. Reporter gene assay demonstrated that E1AD was capable of inhibiting NF-kB activity in the PAEC in-duced by TNF-a without sensitizing to apoptosis, and the rate of inhibition was 53%. Furthermore, E1AD inhibited the expression of a NF-kBdependent inflammatory gene E-selectin in the cells, and the rate of inhibition was 63%. In summary, the usage of E1AD gene may be a new strategy to overcome DXR in the xenotransplantation.
