Influence Of Bortezomib On Acute Myelogenous Leukemia Cells Apoptosis And Its Effects On SALL4Gene And Wnt Signaling Pathway

Objective: To investigate Bortezomib-induced apoptosis of acutemyelogenous leukemia cell lines NB4,TF1and its effect on expressions ofSALL4gene and target genes of Wnt/β-catenin pathway. Methods:Proliferations were analyzed by MTT assay. Apoptosis rates were detectedby flow cytometer. SALL4protein was detected by immunocytochemistry.SALL4proteins in two cell lines were detected by Western Blot. Theexpressions of SALL4,C-myc and CCND1genes in two cell lines weredetected by Real-time PCR. Results: Bortezomib inhibited theproliferations of two cell lines in a time-and-does depending manner.NB4and TF1cells’48h IC50were23.97nmol/L and25.36nmol/L. Bortezomibinduced two cell lines apoptosis in a does-depending manner. After cellswere treated by Bortezomib for24h, the apoptosis rates of NB4cells in30nmol/L and50nmol/L groups were(27.75±1.77)%and (40.45±12.80)%respectively, which had a significant difference in comparing with that ofcontrol group(4.65±3.18)%(P<0.05);50nmol/L group’s apoptosis rate of TF1cells was(64.60±1.31)%,which also had a significant difference incomparing with that of control group(16.15±2.90)%(P<0.05).Immunocytochemistry analysis revealed that both of two cell linesexpressed SALL4proteins which located in cell nucleus. Western Blotrevealed that both of the cell lines expressed SALL4B proteins and whichcould be inhibited by Bortezomib in a time-and-does manner. All the genescould be down-regulated by Bortezomib. There were significantdifferences of gene expressions between experimental group and controlgroup(P<0.05), after cells were treated by50nmol/L Bortezomib for24h.There were significant positive correlations between the alternativeexpressions of SALL4gene and C-myc,CCND1genes. Conclusion:Suppression of SALL4, C-myc and CCND1genes may play a key roleduring Bortezomib-induced NB4and TF1cells apoptosis.