Mechanism Study Of Seleno-Chitosan On Apoptotic Effect In Human Non-small Lung Cancer A549 Cells

The effects of the proliferation inhibition and apoptosis of Human Non-small Lung Cancer Cells A549 induced by Seleno-Chitosan were investigated.The changes of cell morphological,cell cycle,apoptosis pathway and differential protein were explored by MTT,flow cytometry and two-dimensional gel electrophoresis.Firstly,the proliferation inhibition and apoptosis of A549 induced by Seleno-Chitosan were researched.The MTT assays showed that Seleno-Chitosan significantly suppressed the growth of A549 cells in a dose and time dependent manner.The cell growth inhibitory rate of A549 could reach 77%after induced by Seleno-Chitosan(200 μg/mL)for 48 h.The cell morphology had an obviously change.Hoechst33342/PI and DAPI staining revealed that A549 cells induced by Seleno-Chitosan appeared typical apoptotic characteristics such as cell volume reduction,chromatin shrinkage and apoptotic body.The results of Annexin V-FITC/PI dyeing indicated that the apoptosis of A549 cells was gradually from the early apoptosis to late apoptosis after induced by Seleno-Chitosan for different time.FCM analysis demonstrated that the cell cycle was arrested in S phase.Secondly,the apoptosis signaling pathway of A549 cells induced by Seleno-Chitosan was studied.The results showed that the level of Ca2+ and ROS was increase,the MPTP channel was opening,the ΔΨm was decrease.Then,total protein maps were analysed between Normal A549 and A549 cells induced by Seleno-Chitosan for 48 h using SDS-PAGE and two-dimensional gel electrophoresis technology.14 proteins expressed differentially were detected,11 of those were up-expressed and 3 of those were down-expressed.Five proteins were successfully identified by MALDI-TOF-MS/MS.The phospholipase C-alpha,prohibitin iso form 1,were up-expressed,the annexin A4,anterior gradient protein 2 homolog precursor,10 kDa heat shock protein were down-expressed.These five proteins were associated with mitochondrial pathway.In summary,Seleno-Chitosan could effectively inhibit the proliferation of A549 cells and induce apoptosis through the mitochondrial pathway.It was proved that Seleno-Chitosan would be a promising anti-cancer active material in future development.