Gland Of Xage-1b Virus Interference Vector Construction And Function Of A Fiber Deleted Recombinant Adenovirus System To Build

XAGE discovered by bioinformatics, is a new type of cancer/testis antigen (CT antigen), whose main expressing subtype is XAGE-1b and its function and expressing regulation remains unknown. In the first part of this work, we constructed two recombinant adenovirus strains expressing small interfering RNA against XAGE-1b, named Adv-Sh1 and Adv-Sh2. Compared with control cells, semi-quantitative and quantitative RT-PCR showed a higher interfering efficiency of Adv-Sh1 than Adv-Sh2.Cell fluorescence interference assay showed a remarkable efficiency of down-regulation against target gene. Compared with control cells, cells infected with Adv-Sh1 showed higher multiplication rate by cell multiplication assay (P<0.01,Student's t test), and formed more and larger cell colonies by colony formation assay. The result showed down-regulation of XAGE-1b could suppress the multiplication and metastasis of multiple kinds of cancer cells, suggesting an important role of XAGE-1b in tumor growth and metastasis. These findings are significant for further study on XAGE-1b's function and role in tumorigenesis.Adenovirus could not infect a certain kind of cell line specially without infecting others, this is mainly because of the widespread presence of the receptors of adenovirus fiber protein on all kinds of cells, which is a barrier for appliance of adenovirus vector in gene therapy. To knock-out the fiber gene in adenovirus genome and replace it with modified fiber gene is one of the methods of generating targeting adenovirus. In the second part of this work, we designed a new kind of adenovirus targeting system.Based on AdEasy adenovirus system, we constructed a fiber knock-out adenovirus vector. And we also constructed a HEK293 based cell line expressing Fiber protein stably as a packaging cell line for this adenovirus vector. As a result, the fiber knock-out adenovirus vector could be packaged and multiply normally in this cell line. The system would be a significant basis for further construction of adenovirus targeting gene therapy system.