Inhibitory effect of BTEB2 antisense RNA mediated by recombinant adenoviral vector on proliferation of vascular smooth muscle cells / 第三军医大学学报

ABSTRACT

Objective To construct recombinant adenoviral vector expressing basic transcription element binding protein 2 (BTEB2) antisense RNA and study the effects of BTEB2 antisense RNA on the proliferation of vascular smooth muscle cells (VSMCS). Methods BTEB2 cDNA was prepared by RT-PCR technique and was subcloned reversedly into the shuttle plasmid to construct recombinant shuttle plasmid, and then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHG were cotransfected into 293 cells to obtain the recombinant adenovirus. The PCR technique was used to detect the target gene fragment and adenovirus genomic characteristic fragment. After transfection of the recombinant adenovirus into VSMCs, BTEB2 antisense RNA was detected by RT-PCR. The effects of BTEB2 antisense RNA on BTEB2 protein expression, proliferation, and cell cycles of VSMCs were analyzed by Western blot, MTT test, and flow cytometry, respectively. Results The recombinant adenovirus was proved correct by PCR. The expression of BTEB2 antisense RNA was very obvious in VSMCs after infection by recombinant adenovirus. Recombinant adenovirus infection significantly inhibited BTEB2 protein expression and proliferation of VSMCs and resulted in G 0/G 1 block. Conclusion The recombinant adenoviral vector expressing BTEB2 antisense RNA has been constructed. BTEB2 antisense RNA can significantly inhibit the proliferation of VSMCs.