2.Apoptusis of SMMC-7721 cells induced by vitamin K3 via up-regulating Fus expression on the surface of cells
Xiangyang LIN ; Liang SHI ; Yanhui JIN ; Xiaodong CHEN ; Zhihua TAO ; Wenpin CAI
Chinese Journal of Laboratory Medicine 2008;31(7):784-788
Objective To investigate vitamin K3 (VK3) effect on apoptosis of human liver cancer cells and its mechanism. Methods The SMMC-7721 cells were cultured in the experiment. The inhibitory effects of VK3 on SMMC-7721 cells were tested by CCK-8. Morphological evaluation of apoptosis was performed by Hcechst33342 staining. The distribution of cell cycle and apoptosis, and the expression of Fas were assayed by flow cytometry. Fas mRNA expression were detected by RT-PCR. And the concentration of soluble Fas(sFasL) in the culture supernatant were measured by EIJSA. Results The inhibitory rates ofVK3 (at concentrations of 2,5,10,20,25 umol/L for 48 h) on SMMC-7721 growth were 33.8% ,50.1%,63.9% ,78.5% and 84.7%, respectively. Compared with the control group, the number of cells in the G0/G1 phase increased, while that of S phase decreased. The apoptotic cell rates were 18.75%, 25.80%,38.80% ,29.92% and 26.18% ,respectively. The apoptosis cells were strongly stained by Hcechst33342.On exposure to VK3 at the concentration of (2,5,10 umol/L) after 48 h, the mean fluorescence intensity ofFas on cell surface and the expression of fas mRNA and the concentration of FasL in the culture supematantin SMMC-7721 were increased, but they all decreased at the high concentration of VK3. Conclusion VK3can inhibit the proliferation of SMMC-7721 cells and induce apoptosis via up-regulating expression of Fas and sFasL.