Determining the influence of vitamin K3 at different concentrations on hepatoma cells apoptosis cycle by flow cytometry

Objective: To explore the mechanism of vitamin K3-induced hepatoma cells apoptosis so as to guide the clinical treatment of liver cancer.Methods: Logarithmic phase of liver cancer cells were selected from cultured human hepatoma cell line HCC-9204,and were treated with different concentrations of vitamin K3 for different length of time.Apoptotic nuclear morphology was observed under confocal laser scanning microscope.CKK-8 method was used to detect inhibition of HCC-9204 cell proliferation by vitamin K3,flow cytometry was applied to detect apoptosis rate and apoptosis cycle and cell membrane expression of Fas.Results:CCK-8 test showed that hepatoma cell ratio was significantly decreased;growth inhibition rate increased showing an significant measurement dependence.After staining,apoptotic nuclei in metaphase cells presented a corrugated or crease-like shape,parts of the chromatin condensed,late apoptotic nuclear chromatin presented a high degree of cohesion and marginalized,chunky nucleus cracked and produced apoptotic bodies.Flow cytometry showed 48 hours after the interference of VK3 at 2μmol/L leaded to apoptosis rate of 31.44%,5 μmol/L VK3 induced apoptosis rate of 62.81%,10 μmol/L,76.89%;20μmol/L,89.24%;25 μmol/L,93.58%.Conclusions:Vitamin K3-induced hepatoma cell apoptosis shows the dependence of apparent concentration measurement,the fluorescence staining can reveal dynamic changes in the nuclei apoptosis and can be used for both quantitative,qualitative analysis in this study.