Preliminary Study of Seleno-chitosan-induced Apoptosis in A549 Cells through a Mitochondrial Signaling Pathway
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Abstract:
The effects of seleno-chitosan (SC) on cell proliferation inhibition and apoptosis in human non-small-cell lung cancer A549 cells were investigated, and the differential expression of proteins in SC-treated and untreated cells were determined. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was employed to determine the inhibitory effect of SC on the proliferation of A549 cells. Morphological changes in the A549 cells after Hoechst 33342/PI staining and the opening of the mitochondrial permeability transition pore (MPTP) after calcein staining were observed by laser scanning confocal fluorescence microscopy. The effect of SC on mitochondrial membrane potential (ΔΨm) was analyzed by flow cytometry (FCM). Furthermore, two-dimensional gel electrophoresis (2-DE) combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) was used to identify the differential expression of apoptosis-related proteins. The results showed that SC could significantly suppress the proliferation of A549 cells in a time- and concentration-dependent manner. Morphological changes of A549 cells could be induced by SC; the cell shape became irregular and chromatin condensation and margination occurred. After SC treatment, the MPTP of A549 cells was gradually opened. Additionally, two differentially expressed proteins, prohibitin isoform 1 and heat shock protein, were identified by MALDI-TOF.
