The Influence Of Adenovirus Ad-p55PIK-GFP On Tumor Formation Of Cancer Cell

Background and objective:Phosphoinositide 3-kinases family play an important role in numerous cell signaling transduction pathways, including cell proliferation, cell transformation, cell apoptosis and cell differentiation. Abnormal activation of PI3Ks is observed in most of human cancer. p55PIK, a regulatory subunit of PI3Ks, may be functional as an important regulator in cancer proliferation. The object of study is to construct recombinant adenovirus carrying p55PIK gene and observe the effect of p55PIK on tumor formation in vivo.Methods:Utilize Flag-cmv-p55PIK plasmid as a template, p55PIK gene was amplified by PCR, then been construction into the shuttle plasmid pAdTrack-cmv. After that, the construction plasmid pAdTrack-CMV-p55PIK is disgested by PmeI, then cotransfected with bone plasmid pAdEasy-1 into germ of BJ5183, identified by enzyme digestion. Then the Ad-p55PIK-GFP is digested by PacI and transfected into the HEK293A cell. By observing the green fluorescence, the recombinant adenovirus Ad-p55PIK-GFP is holded in HEK293A cell. After amplified for three circles, a large number of adenovirus is harvested. After that, the recombinant adenovirus Ad-p55PIK-GFP is transfected into SW480 colon cell, then plant the SW480 cell in the subcutaneous of Nude, observing the effect on tumor cells into nude miceResults:The result of enzyme digestion means that the recombinant adenovirus plamid of pAd-p55PIK-GFP is successfully constructed. The green fluorescences mean that the Ad-p55PIK-GFP is holded in HEK393A cell. After transfected into the SW480 cell, the high expressing of protein of p55PIK is more increased than the untransfected. The tumor transfected by Ad-p55PIK-GFP are larger than the control group and the empty Ad group.Conclusion:The recombinant adenovirus Ad-p55PIK-GFP is successfully constructed. The high expression of p55PIK in tumor cells more conducive to the tumor formation in vivo.