Effects of dead box 1 gene on invasion, migration and drug resistance capability of neuroblastoma cells

Abstract

Objective To explore the effects of dead box 1(DDX1) gene on invasion, migration and drug resistance capability of neuroblastoma(NB) cells. Methods According to the virus drop degree, the appropriate amount of target virus(Lenti-DDX1-MIR virus liquid, drop degrees 1012 TU/L) and negative control virus(Lenti-EGFP virus liquid, drop degrees 3×1011 TU/L)(multiplicity of infection was 10) were added into 2 hole cells, respectively.SK-N-BE(2)/blank, SK-N-BE(2)/shV and SK-N-BE(2)/shDDX1 cells which grew in good condition were cultured.Transwell chamber was used to detect the invasion, and cell staining was made with crystal violet.The researchers calculate 5 field counting in each small room and calculate the average cell invasion rate.Transwell chamber was used to detect the migration, and cell staining was made with crystal violet.The 570 nm absorbance values was tested with enzyme linked immunosorbent assay(ELISA) reader, and cell migration was calculated.The researchers used 50 mg Cisplatin, solution 10 g/L mother liquor standby with 5 mL dimethyl sulphoxide, and 50 mg Doxorubicin, solution 1 g/L mother liquor standby with PBS.Drugs were added to the cell culture plate, and Doxorubicin final concentration was 1.0 mg/L, and Cisplatin final concentration was 2.5 mg/L, and photographic record was documented after drug treatment for 24 h. Cell Count Kit-8(CCK-8) was used to detect the drug sensitivity to NB cells to Doxorubicin and Cisplatin. Results Transwell results showed that, cell invasion concentration in SK-N-BE(2)/shDDX1 was 60% compared with SK-N-BE(2)/blank and SK-N-BE(2)/shV; Crystal violet staining showed that cell invasion of SK-N-BE(2)/shDDX1 was significantly weaker than that of SK-N-BE(2)/blank and SK-N-BE(2)/shV cells, that is to say, DDX1 knockdown reduced the cell invasion of NB.Transwell results showed that, cell migration concentration in SK-N-BE(2)/shDDX1 was 50% compared with SK-N-BE(2)/blank and SK-N-BE(2)/shV; Crystal violet staining showed that cell migration of SK-N-BE(2)/shDDX1 was significantly weaker than that of SK-N-BE(2)/blank and SK-N-BE(2)/shV cells, that is to say, DDX1 knockdown reduced the cell migration of NB.With DDX1 knockdown, 24-h inhibition rate of SK-N-BE(2)/shDDX1 cell was 1.93 times of SK-N-BE(2)/shV cell with 1.0 mg/L Doxorubicin, 24 h inhibition rate of SK-N-BE(2)/shDDX1 cell was 1.38 times of SK-N-BE(2)/shV cell with 2.5 mg/L Cisplatin.DDX1 knockdown could increase the Doxorubicin and Cisplatin drug sensitivity to NB cells. Conclusion DDX1 knockdown can decrease the cell invasion, migration and resistance capability of NB and increase the Doxorubicin and Cisplatin drug sensitivity of NB cells. Key words: Dead box 1 gene; Neuroblastoma; Invasion; Migration; Drug resistance