Objective    To construct the recombinant co-expression vector carrying antisense RNAs to dual-target BAG-1 and Bcl-2 and investigate its effect on the proliferation of SGC-7901 cells.     Methods    RT-PCR was used to amplify the full length CDS of BAG-1 and Bcl-2 cDNA from total RNA of SGC-7901 cells. The cDNA fragments of BAG-1 and Bcl-2 were inserted into eukaryotic co-expression pVITRO2 vector multiple cloning sites in the antisense orientation respectively by gene recombination technology. Then the recombinant plasmid was transfected into SGC-7901 cell. The proliferation of the transfected SGC-7901 cells was determined by MTT assay. The changes of BAG-1 and Bcl-2 mRNA levels were detected by RT-PCR and the division of cell cycle was detected by FCM.     Results    The restriction endonucleases digestion and sequencing suggested that the eukaryotic co-expression vector pVITRO2-AsBAG-1-Bcl-2 was constructed successfully. MTT assay demonstrated that pVITRO2-AsBAG-1-Bcl-2 could inhibit the proliferation of SGC-7901 cells in a time-dependent manner

with most significant effect in 72 h after transfection (P0.01). BAG-1 and Bcl-2 mRNA expressions in pVITRO2-AsBAG-1-Bcl-2 transfected cells were significantly decreased compared with those in the control cells and pVITRO2 transfected cells (P0.05). The apoptotic rate of pVITRO2-AsBAG-1-Bcl-2 transfected cells was significantly higher than that in control cells and pVITRO2 transfected cells (P0.01).     Conclusion    The co-expression vector pVITRO2-AsBAG-1-Bcl-2 has been successfully constructed and it can inhibit the proliferation of SGC-7901 cells and induce cell apoptosis.