Effects Of Diallyl Disulfide On Differential Expression Of Apoptosis Associated Genes In Leukemia Cell Line K562

ObjectiveTo investigate the differentially expressed profile of apoptotic gene in K562cellsinduced by Diallyl disulfide(DADS).Methods1. The morphologic changes of leukemia K562cells were observed byWright-Gimsa staining and Hoechst33342staining after24h,48h,72h of40mg/L DADStreatment.2. The expression profile of apoptosis-associated genes between K562cells beforeand after treated with DADS, was identified by gene array.3. The up-regulated GADD45A and down-regulated HO-1gene were confirmed byreverse transcription-polymerase chain reaction (RT-PCR).Results1. Wright-Gimsa staining finds the typical morphologic changes of apoptosis after40mg/L DADS treated K562cells: cell volume reduction, shape irregular, the ratio ofnuclear and cytoplasm decreasing, even sprouting.2. Hoechst33342staining finds the typical morphologic changes of apoptosis after40mg/L DADS treated K562cells: cell nucleolus reducion, chromatin condensation,chromatin debris, even apoptotic body.3. When treated with40mg/L DADS for24h, the differential expression of369genes, and14apoptosis-associated genes were found with gene array,5of which wereup-regulated gene: GADD45A, HSPA1B, PACAP, AEN, TPT1, and9weredown-regulated: CDKN2D, NFKBIA, IL7, PPP1R13L, GCLM, DDLT4, HO-1, BNIP3L,BNIP3. 4. RT-PCR results exhibited that a increased GADD45A mRNA expressioncompared with the control group and a decreased HO-1mRNA expression after40mg/LDADS treated K562cells for24h, were consistent with the detected by gene array.Conclusion1. The apoptosis be mediated by some specific genes after40mg/LDADS treatedK562cells for24h.2. The signal pathways, which is related with GADD45A and HO-1genes, canregulate apoptosis of DADS inducing K562cells.