Construction and application of recombinant adenovirus vector expressing the mouse endostatin

Objective To construct the recombinant adenovirus vector expressing the mouse en-dostatin and investigate where endostatin expressed by the infected pancreatic carcinoma cells(BxPC-3, AsPC-1)can inhibit angiogenesis. adenovirus. Methods The mEndostatin DNA extracted from vector containing mouse endostatin gene was successfully amplified using PCR and cloned into adenovirus vector pCA13,named pCA-mEndo. Recombinant adenovirus plasmid pCA13-mEndo was cotransfected with pBHG3 into 293 packaging cells by lipofectamine 2 000 and the replication-deficient recombinant adenovirus Ad-mEndo recombinant adenovirus was purified by CsCl density purification and the titer measured pancreatic adenocarcinoma cells(BxPC-3, AsPC-1)were infected with adenovirus Ad-mEndo in vitro. Supernatants were measured and the expression of endostatin proteinwas analyzed for their ability to inhibit angiogenesis. Results The mEndostatin DNA was confirmed by sequence meusure-ment. The virus titer was 6. 3 × 1010 pfu/ml. Ad-mEndo can efficiently infected pancrealic carcinoma cells and the expressed mEndoslatin can inhbit angiogenesis. Conclusion The conslrucled adenovirus Ad-mEndo can efficienlly express mEndoslalin, which can inhibit angiogenesis. This invesligation provides the basis for sludying pancrealic carcinoma gene therapy in vivo.