Apoptosis in Jurkat Cells Induced by Lfcin from Different Sources
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Abstract:
In this study, the inhibitory effects of bovine lactoferricin (LfcinB) and human lactoferricin (LfcinH) on the proliferation of Jurkat cells were explored and compared, by reducing the mitochondrial membrane potential in the cells. The effect on Jurkat cell proliferation was measured by MTT assay, and nuclear changes were observed by fluorescence microscopy with Hoechst 33258 staining. A flow cytometry of double-labeling by Annexin V-FITC/PI was used to distinguish the apoptosis stages in the Jurkat cells that were induced by LfcinH and LfcinB. The results showed that LfcinB and LfcinH significantly inhibited the proliferation of Jurkat cells (P < 0.05) in a dose-dependent manner. A decreased mitochondrial membrane potential in Jurkat cells was observed by JC-1 staining using laser scanning confocal microscopy. The early phase of apoptosis occurred after Jurkat cells were treated by LfcinH and LfcinB for 48 h; characteristics of apoptosis such as nuclear shrinkage and debris were observed using fluorescence microscopy. The results suggest that the change in mitochondrial membrane potential may have led to apoptosis in Jurkat cells and, eventually, the inhibition of proliferation. When the concentration of both, LfcinH and LfcinB was less than 250 μg/mL, the inhibitory effect of LfcinH on Jurkat cells was noted to be stronger; whereas at concentrations higher than 250 μg/ mL, LfcinH and LfcinB showed similar inhibitory effects.
