Effects of dead box 1 gene on cell apoptosis, proliferation and cell cycle of neuroblastoma cells

Abstract

Objective To explore the effects of dead box 1(DDX1)gene on cell apoptosis, proliferation and cell cycle of neuroblastoma(NB) cells. Methods SK-N-BE(2)/blank, SK-N-BE(2)/shV and SK-N-BE(2)/shDDX1 cells were seeded in 96-well plates, which grew in good condition and in the logarithmic growth phase, 5 000 cells were inoculated in each well and 5 repeated holes were set.Cell count kit 8(CCK-8) was used to detect the cell number at 12 h, 24 h, 36 h, 48 h, and the average was calculated.The time (hour) was set as abscissa, the optimal density (A) value at 450 nm was set as vertical axis, and the growth curves of these 3 cells were drawn to investigate the effects of DDX1 on the proliferation of NB cells.After 24 h, flow cytometry (PI staining) was used to detect the apoptosis of SK-N-BE(2)/blank, SK-N-BE(2)/shV and SK-N-BE(2)/shDDX1 cell lines to observe the effects of DDX1 on the apoptosis of NB cells.After 24 h, flow cytometry (PI staining) was used to detect the proportion of SK-N-BE(2)/blank, SK-N-BE(2)/shV and SK-N-BE(2)/shDDX1 cells at G1, S, M, G2 stage to observe the effects of DDX1 on the cell cycle of NB cells. Results SK-N-BE(2)/shDDX1 cell proliferation was significantly lower than SK-N-BE(2)/blank and SK-N-BE(2)/shV cells, that was to say, DDX1 knockdown reduced the cell proliferation of NB.According to the flow cytometry results, the total average apoptosis rate was 5.28% in SK-N-BE(2)/shV cells, and the total average apoptosis rate was 9.99% in SK-N-BE(2)/shDDX1 cells.The number of apoptotic SK-N-BE(2)/shDDX1 cells was significantly higher than the number of SK-N-BE(2)/blank and SK-N-BE(2)/shV cells, which indicated that DDX1 knockdown increased tumor cell apoptosis of NB.Compared with SK-N-BE(2)/blank and SK-N-BE(2)/shV cells, the cell cycle of SK-N-BE(2)/shDDX1 cells was arrested, and the proliferation was affected. Conclusions After DDX1 expression is inhibited, the cell cycle of NB cells are affected, the cell apoptosis is increased, and the cell proliferation is reduced. Key words: dead box1 gene; Neuroblastoma; Cell cycle; Apoptosis; Proliferation