[Effect of overexpression of CAV1 mediated by lentivirus on proliferation and apoptosis of HL-60 cells].

This study was purposed to explore the effect of lentivirus-mediated CAV1 overexpression on proliferation and apoptosis in HL-60 cells.Recombinant lentiviral expression vector pcDNA-EF1-CAV1 was constructed,and cotransfected the 293TN cells with a mixture of pPACK packaging plasmids.Then collecting virus suspension infects the HL-60 cells,which make CAV1 gene stable transfection and high expression in the cells.The CAV1 protein expression status in HL-60 cells transfected was elvaluated through Western blot method.Proliferative activity and apoptosis of HL-60 cells before and after transfection were detected by CCK-8 method and flow cytometry,respectively.The results showed that the PCR-positive clone screening and results of nucleotide sequencing confirmed that the CAV1 gene inserted into the expression vector pcDNA-EF1-GFP correctly,recombinant lentiviral particles Lv-CAV1 transfected HL-60 cells successfully and with transfection rate up to 90%.The result of Western blot showed that CAV1 protein expression in HL-60 cells significantly increased at 48 hours after transfection.CCK-8 result indicated that cell proliferation activity increased at 48 h after transfection(P0.05),flow cytometry testing results displayed that apoptosis rate of HL-60 cells obvirously decreased after transfection(P0.01).It is concluded that the overexpression of CAV1 in HL-60 cells can inhibit cell proliferation activity and promote cell apoptosis.