Objective: To explore the effect of autocrine vascular endothelial growth factor (VEGF) on the abnormal proliferation of HL-60 cells.
Methods: HL-60 cells were transfected via lipofictamine with the VEGF(165) cDNA sense vector (HL-60/ VEGF(165)) and with the PcDNA3.1(-)-vector (HL-60/neo) as the control. Reverse transcription-polymerase chain reaction (RT-PCR) was used for detecting VEGF mRNA in the transfected cells. VEGF concentrations in the cell culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA), and cell proliferation was determined by MTT assay and colony-forming assay in vitro. Flow cytometric Annexin-V-FITC/ PI dual-labeling technique was employed to observe the effect of VEGF(165) cDNA transfection on the apoptosis of HL-60 cells.
Results: The levels of VEGF mRNA expression by HL-60/VEGF(165) cells were higher than HL-60/neo, with the mean VEGF quantity in the supernatant of HL-60/VEGF(165) cell culture being 399.07+/-12.45 ng/L, which was 2-fold higher than that in the supernatant of HL-60/neo cells (184.45+/-10.53 ng/L) (P<0.01). The growth rate of HL-60/VEGF(165) cells was significantly accelerated as compared with that of the control cells, and the colony formation capacity of the former cells also increased significantly (P<0.05), with the average colony number of 157.00+/-17.00/500 cells vs 110.00+/-12.90/500 cells of HL-60/neo cells. Less apoptotic cells were identified among HL-60/VEGF(165) cells than in HL-60/neo cells in the same culture condition.
Conclusion: Autocrine VEGF plays an important role in the proliferation and apoptosis of acute myeloid leukemia cells.