BACKGROUND AND OBJECTIVE: Diallyl disulfide (DADS), an antitumor reagent, has increasingly gained attention. This study was to explore the related proteins in DADS-induced apoptosis initiation in human leukemia HL-60 cells. METHODS: Total protein of HL-60 cells with or without 2-day treatment of 3.6 mg/L DADS was extracted, separated by two-dimensional polyacrylamide gel electrophoresis, and analyzed by PDQuest 2-DE software. Differentially expressed proteins were separated and identified by peptide mass fingerprinting analysis and bioinformatics, and verified by western blot and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: As compared with those in untreated HL-60 cells, 29 proteins were differentially expressed in DADS-treated HL-60 cells: 22 were upregulated and seven were downregulated. Among nine proteins which were randomly selected for peptide mass fingerprinting analysis and bioinformatics, seven were meaningful. These proteins were associated with cellular responses, gene transcription and regulation, cytoskeleton, metabolism, and so on. western blot showed that Ras-related C3 botulinum toxin substrate 2 (Rac2) was upregulated in DADS-treated HL-60 cells; this result was accordant with RT-PCR results. CONCLUSION: Some proteins, including Rac2, may be involved in DADS-induced apoptosis in human leukemia HL-60 cells, but the exact mechanism needs to be explored.
BACKGROUND AND OBJECTIVE:Diallyl disulfide (DADS), an antitumor reagent, has increasingly gained attention. This study was to explore the related proteins in DADS-induced apoptosis initiation in humanleukemia HL-60 cells. METHODS: Total protein of HL-60 cells with or without 2-day treatment of 3.6 mg/L DADS was extracted, separated by two-dimensional polyacrylamide gel electrophoresis, and analyzed by PDQuest 2-DE software. Differentially expressed proteins were separated and identified by peptide mass fingerprinting analysis and bioinformatics, and verified by western blot and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: As compared with those in untreated HL-60 cells, 29 proteins were differentially expressed in DADS-treated HL-60 cells: 22 were upregulated and seven were downregulated. Among nine proteins which were randomly selected for peptide mass fingerprinting analysis and bioinformatics, seven were meaningful. These proteins were associated with cellular responses, gene transcription and regulation, cytoskeleton, metabolism, and so on. western blot showed that Ras-related C3 botulinum toxin substrate 2 (Rac2) was upregulated in DADS-treated HL-60 cells; this result was accordant with RT-PCR results. CONCLUSION: Some proteins, including Rac2, may be involved in DADS-induced apoptosis in humanleukemia HL-60 cells, but the exact mechanism needs to be explored.