title: [Effect of overexpression of CAV1 mediated by lentivirus on proliferation and apoptosis of HL-60 cells].
abstract: 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 evaluated 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 (P < 0.05), flow cytometry testing results displayed that apoptosis rate of HL-60 cells obviously decreased after transfection (P < 0.01). It is concluded that the overexpression of CAV1 in HL-60 cells can inhibit cell proliferation activity and promote cell apoptosis.
authors.affiliation: Biochemical Laboratory, Basic Medical College, Jiamusi University, Jiamusi 154000, Heilongjiang Province, China; Office of Master Postgraduate, Mudanjiang Medical College, Mudanjiang 157011, Heilongjiang Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ma
authors.forename: Di-Di
authors.initials: DD
authors.lastname: Wang
authors.forename: Zhao
authors.initials: Z
authors.lastname: Wang
authors.forename: Gui-Ming
authors.initials: GM
authors.lastname: Zhu
authors.forename: Peng-Xia
authors.initials: PX
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism*; D049109:Cell Proliferation; D005822:Genetic Vectors; D018922:HL-60 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D010957:Plasmids; D014162:Transfection
chemical_list: C495540:CAV1 protein, human; D051242:Caveolin 1