Kistrin Induced Cell Apoptosis And Cell Cycle Change In Human Lens Epithelial Cells Line

Objects:To investigate the effects of kistrin on cell apoptosis and cell cycle in human lens epithelial cells lines SRA01/04.Methods:All the cultured SRA01/04 cell the study used were in the logarithmic growth phase.â‘ .Apoptosis Assay:SRA01/04 cell were cultured with DMEM/F12 medium containing 10%fetal bovine serum.They are incubate with medium containing different concentration of kistrin(0ng/ml,80ng/ml,160 ng/ml,320 ng/ml)for 8,16 and 32 hour.The first group was control group.And then,apoptosis was detected by flow cytometer(FCM).â‘¡Morphology change on cell apoptosis:At the logarithmic growth phase,cells are incubate with DMEM/F12 medium containing different concentration of kistrin (0ng/ml,160ng/ml)for 24 and32 hours.Then stained by acridine and observed by Fluorescence Inverted Microscope.We take photos before staining.â‘¢Cell cycle assays by flow cytometry:the SRA01/04 cell were seeded in 6-well plate by using DMEM/F12 medium and containing 10%fetal bovine serum.After 48hours,the medium is changed to serum-free media for synchronization about24h.And then the medium is changed to DMEM/F12 medium containing 10%fetal bovine serum,with kistrin at the concentration of 0ng/ml,160 ng/ml and 320ng/ml.Cell cycle is assays by flow cytomete 24h later.Resulits:1.Kistrin can induce apoptosis significantly.At the 8hours,the apoptosis rates of kistrin at concentrtion of 0ng/ml,80ng/ml,160 ng/ml,320 ng/m are 1.29%,5.48%,11.23%,20.77%;at the 16hours,the apoptosis rates of kistrin at concentration of 0ng/ml,80ng/ml,160 ng/ml,320 ng/ml are 3.04%, 7.68%,19.76%,32.36%.The apoptosis rates of kistrin at concentration of 0ng/ml,80ng/ml,160 ng/ml,320 ng/m come to the 4.75%,72.15%,54.09%, 43.27%when it was 32hours.There are significant difference between control group and experimental group.The apoptosis rates ascend with the increasing of kistrin concentration.2.There is a significant morphology change when the cells are incubate with kistrin(160 ng/ml)for 24 hour,but the apoptosis can not be oberserve by acridine stained.32h later,the apoptosis can be found,but not the necrosis.3 Cell cycle is block by using kistrin at the concentrantion of 160ng/ml and 320ng/ml,24 hours latter.Percentage of G1 about control group is 36.9%,much lower then the experimental group 66.0%(160ng/ml)and 69.3%(320ng/ml).Cell cycle is block significantly at G1 phase.Conclusion:A disintegrin-kistrin,has the ability of inducing apoptosis,it can inhibit the cell proliferative by blocking cell cycle at the G1 phase.