Aim To construct short hairpin RNA(shRNA)retroviral vector of PKB/Akt and study the effect on the proliferation of vascular smooth muscle cell(VSMC).Methods The sequence of shRNA of PKB/Akt was constructed by the software and synthesized by chemical method and then was cloned into pGEM-T vector,which was certified by DNA sequencing.The sequence was inserted into a retroviral vector pLXIN,then the vector was packaged in PT67 cells.The efficiency of inhibition was verified by Northern blot and Western blot after transfection to VSMC.The change of p70s6k was also determined at the same time.The proliferation activity was determined by flowcytometry and MTT.Results The PKB/Akt shRNA vector was successfully constructed by means of recombinant DNA technology and cell transfection.Additionally,the vector could efficiently decrease mRNA and protein expression of PKB/Akt,while the expression of p70s6k also decreased significantly.The cell cycle VSMC was also stunned in phase G0/G1.Conclusion The PKB/Akt shRNA retroviral vector has been constructed successfully,which can significantly inhibit the proliferation of VSMC.