A Preliminary Study On The Vitamin K2 Induced Hepatocellular Carcinoma Cells Autophagy

Objective:The purpose of this study was to explore the potential inhibitory effects of vitamin K2(Vitamin K2,VK2)on Hepatocellular carcinoma cells and to explore the possible mechanisms.Methods:Bel-7402 cells and Hep G2 cells were cultured and amplified in vitro.Then they were pre-treated as originally planned.The CCK-8 was used to study the proliferation;flow cytometry was used to detect cell cycle and cell apoptosis of Bel-7402 cells.Western Blot was used to test the proteins about apoptosis.Western Blot was used to test the protein about autophagy by add the chloroquine.Results:In this study,we found VK2 can significantly suppress cell proliferation of Bel-7402 cells and Hep G2 cells.Cell cycle distribution analysis showed that cells of combination group were blocked in the G1 phase.Apoptosis analysis shows that the apoptosis rate of control group,50?M VK2 group,200?M VK2 group(all cultured for 48hours)was 0.28±0.038%,0.62±0.27% and 1.23±0.69%(p>0.05)respectively.The result of Western Blot showed that VK2 could up regulated Cleaved C cytometry aspase 3 and Cleaved PARP in Bel-7402 cells.VK2 could up regulated LC3-??LC3-?/LC3-?(p<0.01)?PINK1(p<0.01)?p62/SQSTM1(p<0.01)?Parkin(p<0.05).When add chloroquine to inhibited autophagy,LC3-??LC3-?/LC3-?(p<0.05)?PINK1(p<0.01)?and p62/SQSTM1(p<0.05)were more up regulated,p-mTOR(p<0.001)and Parkin(p<0.05)was down regulated.Conclusion:This study had shown that VK2 inhibited growth of Bel-7402 cells and Hep G2 cells in vitro and altered in a time-and dose-dependent manner in Bel-7402 cells.Apoptosis was detected by flow and Western Blot was used to detect apoptosis-related and autophagy-related proteins,suggesting that VK2 inhibits growth of Bel-7402 cells in vitro and may be related to autophagy.VK2 has the potential for adjuvant treatment of liver cancer.