Title: Recombinant vector carrying antisense RNA to dual-target BAG-1 and Bcl-2 induces SGC-7901 cell apoptosis
Abstract: 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.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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