Preparation of human vector and its anti-growth effect in the prostate cancer cells α-tocopherol associated protein recombinant adeno-associated virus type 2

Objective To produce recombinant rAAV2 virus carrying α-tocopherol associated protein (TAP) and to observe its effect on prostate cancer cells. Methods The pSNAV-TAP-EGFP plasmid was constructed and digested with restriction enzyme. Polymerase chain reaction (PCR), and sequence analysis were performed to identify the correct recombinant clones. The plasmid was transfected into 293T cells to produce virus. The virus was amplified and purified. DU-145 cells were infected by rAAV-TAPEGFP. Cell growth was detected by MTT assay. Results The pSNAV-TAP-EGFP was constructed successfully. The titer of rAAV2-TAP-virus was 6. 4 × 1011 vg/ml and the purity was 95%. The rAAV-TAP-EGFP was infected into the DU-145 cells with high efficiency. The expression of TAP was increased significantly by real-time PCR. MTT assay showed differences at the 4th day. The cellular absorbance value in the TAP transfection group, control group and empty vector transfection group was 0. 546 +0. 072, O. 673 +0. 015,and 0. 638 + 0. 051 (P ＜ 0. 05 ). The differences became more significant after 6 days. Conclusion High tier recombinant adeno-associated virus rAAV2-TAP-EGFP was obtained, which was able to inhibit the proliferation of prostate cancer cells. Key words: α-tocopberol associated protein; Recombinant adeno-associated virus;  Prostate carcinoma;  Proliferation