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OBJECTIVE:To explore the effect of the specific inhibitor-LY294002 of the signaling pathway PI3K/AKT on cell cycle and apoptosis of HL60/VCR cells and its mechanism.METHODS:Vincristine(VCR) resistant HL60/VCR were treated with LY294002(final concentration at 1.0,2.5,5.0,or 10 ?mol?L-1) or without LY294002(control).IC50 was detected by MTT;flow cytometry was used to evaluate apoptosis and cell cycle distribution,and changes of Bcl-2,CyclinD1 gene mRNA levels were detected by reverse transcription polymerase chain reaction.RESULTS:As compared with control,LY294002 significantly reduced the IC50 value in HL60/VCR to vincristine in a concentration dependent manner(P
Résumé
Objective To explore the effect of vascular endothelial growth factor antisense oligonucleotides(AS-VEGF)on HL60 cell and HL60/VCR multidrug resistance cell and analyze the function of P-gp and the expression of related multidrug resistance genes including Bcl-2,Mcl-1,MDR1,MRP,GST?,TopoⅡ? and TopoⅡ?.Methods A vector AS-VEGF which expressed in eukaryotic cell was established,then transfected the vector into HL60 and HL60/VCR by limposome transfection technology,observed and drew the growth curve by Tapanlan taining,RT-PCR was used to detect the expression of Bcl-2,Mcl-1,MDR1,MRP,GST?,TopoⅡ? and TopoⅡ? in mRNA level after transfected 24 h and 48 h.Western Blot was used to detect the expression of P-gp in proteinum level after transfected 24 h and 48 h.Results The growth of HL60 and HL60/VCR was inhibited by AS-VEGF(1.25 mmol/L).Between HL60 and HL60/VCR,AS-VEGF decreased the expression of MDR1,MRP,GST? and TopoⅡ? but could not influence the expression of Bcl-2,Mcl-1 and TopoⅡ?,and the expression of P-gp was also obviously decreased in 48 h compared with that in 24 h.Conclusions AS-VEGF can inhibite the growth of HL60 and HL60/VCR and reverse multidrug resistance by changing cell microenvironment and the cell membrane correlated protein transportating channel,reduce the cell disintoxicating and the self-repair ability.