Effect of overexpression of CAV1 mediated by lentivirus on proliferation and apoptosis of HL-60 cells.
10.7534/j.issn.1009-2137.2013.04.018
- Author:
Wei MA
1
,
2
;
Di-Di WANG
;
Zhao WANG
;
Gui-Ming ZHU
;
Peng-Xia ZHANG
Author Information
1. Biochemical Laboratory, Basic Medical College, Jiamusi University, Jiamusi 154000, Heilongjiang Province, China
2. Office of Master Postgraduate, Mudanjiang Medical College, Mudanjiang 157011, Heilongjiang Province, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Caveolin 1;
genetics;
metabolism;
Cell Proliferation;
Genetic Vectors;
HL-60 Cells;
Humans;
Lentivirus;
genetics;
Plasmids;
Transfection
- From:
Journal of Experimental Hematology
2013;21(4):905-910
- CountryChina
- Language:Chinese
-
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.
