Influence Of The Recombinant Adenovirus Vector Carrying Apoptin And Endostatin Genes On The Growth Of Subcutaneous Gastric Cancer In Mice

Objective To study the effect of adenovirus vector carrying apoptin and endostatin genes on the growth of subcutaneous gastric cancer in mice.Methods The recombinant adenovirus containing endostatin gene was constructed. The genes of endostatin were amplified by PCR, and the recombination shuttle vector pDC316-Endostatin-mCMV-EGFP was constructed. After co-transfection of the shuttle vector pDC316-Endostatin-mCMV-EGFP and framework plasmid into 293 cells, the recombinant adenovirus carrying endostatin gene was obtained through homologous recombination.The endostatin gene in advenovirus genome was detected by PCR, and the titer of adenoviral vector was detected. A model of subcutaneous gastric cancer in mice was established with mouse fore-stomach carcinoma(MFC), and the mice were randomly divided into groups A,B,C,D and E. The mice of group A was injected with adenovirus vector carrying apoptin and endostain genes, the mice of group B with adenovirus carrying apoptin genes, the mice of group C with adenovirus carrying endostatin genes, the mice of group D with adenovirus carrying EGFP genes, and the mice of group E with PBS buffer. During 12 days after treatment, the volume of the tumors and the adverse effect were observed. All mice were sacrificed and the inhibition rate was calculated. The expression of apoptin and endostatin genes was tested by RT-PCR and the in-situ apoptosis apoptosis of tumor cell was tested by TUNEL.Results Resultant shuttle plasmid was identified by restrictive enzyme digestion and its sequence was assayed. After co-transfection of the shuttle vector pDC316-Endostatin-mCMV-EGFP and framework plasmid into 293 cells,the recombinant adenovirus carrying endostatin genes was successful constructed. PCR test indicated that endostatin gene was successfully integrated into the adenovirus genome, and the titer of adenovirus vector was 1.26×1010 (IU/ml). The volume of tumors in group A was significantly smaller than other groups(P<0.05), while the inhibition to tumor and the apoptotic index were higher than other 4 groups. The apoptin and endostatin genes were expressed in group A, moreover apoptin gene was expressed in group B and endostatin gene was expressed in group C.Conclusion It is concluded that the recombinant adenovirus vector carrying endostatin gene can be successfully packaged, the recombinant adenovirus carrying apoptin and endostatin genes can inhibit the growth of the gastric cancer and induce the apoptosis in mice.