Effect of MEK-ERK signaling pathway on diallyl disulfide-induced autophagy in human leukemia K562 cells / 白血病·淋巴瘤

ABSTRACT

Objective To study the effect of MEK-ERK signaling pathway on diallyl disulfide (DADS)-induced autophagy in human leukemia K562 cells. Methods K562 cells were divided into experimental group, solvent group and blank control group. K562 cells in the experimental group were cultured with 10, 20, 40, 80 mg/L DADS for 48 h. The morphological changes were observed by inverted microscope, monodansyl cadaverine (MDC) staining was used for detecting the formation of the autophagic vacuoles (AV), and the rates of autophagy were analyzed by flow cytometry. Western blot was used to determine the level of expression of ERK, phospho-ERK (p-ERK), LC3-Ⅱ. Results With the increasing of DADS concentration, the number of K562 cells was decreased significantly, and the morphology of some K562 cells became irregular and membrane deformed.The staining of cells and the number of green spots in the cells increased, suggesting that autophagy of the K562 cells cultured with DADS increased. The autophagy rates of K562 cells increased gradually after cultured with DADS for 48 h, the autophagy rate in 40 mg/L DADS group was the highest, and the autophagy rates in 20, 40 and 80 mg/L DADS group were higher than that in the blank control group (P<0.05). There were no significant difference in the expression level of ERK protein between each group (P> 0.05), with the increasing of DADS concentration, the protein expression of p-ERK and LC3-Ⅱ increased, there was significant difference in the protein expression of 40 mg/L DADS group (P<0.05). Conclusion DADS may induce autophagy of K562 cells by activating MEK-ERK signaling pathway through up-regulation of ERK phosphorylation.