Effects Of Supernatant Of Hepatocellular Carcinoma Cells On The Activity Of Echinococcus Granulosus Protoscoleces In Vitro Culture

Objective:To compare the effects of the three hepatocellular carcinoma cells supernatants on the activity of the protoscoleces of Echinococcus granulosus in vitro.Methods:Serum-free supernatants of three tumor cells,Huh7,Hepa1-6,and HepG-2,were collected after tumor cells cultivated 48 h,then co-cultured with the protoscoleces of Echinococcus granulosus（about5,000 in each group）.The control group was cultured in serum-free medium,only.On the 1st,3rd,5th,and7th days after culture,50μl of the suspension was taken for trypan blue staining in each group to observing,counting,and calculating the survival rate.On the 7th day after culture,the microscopic morphology of the protoscoleces wree observed under a scanning electron microscope.And ELISA was used to determine the activity of Caspase-3 in four groups of Echinococcus granulosus.Results:Trypan blue staining showed that the survival rate of the protoscoleces of the serum-free group,Huh7 group,Hepa1-6 group,and HepG-2 group decreased significantly on the 5th day,respectively:（46.54%±1.11%）,（46.19%±3.60%）,（55.90%±2.33%）and（68.73%±1.06%）.On the 7th day,the protoscoleces survival rates of Hepa1-6 and HepG2 groups were 36.09%±0.95%and 45.69%±0.40%,respectively,which were significantly different from the control group at 17.99%±1.52%（P<0.01）.The Huh7 group at 19.04%±1.70%was not significantly different from the control group at17.99%±1.52%（P>0.05）.On the 7th day,scanning electron microscopy showed that the morphology of the protoscoleces of the control group and Huh7 group was generally destroyed,and the ultrastructure of microhairs and suction cups was disordered and disintegrated.The structure is also clear and complete.The cysteine protease-3（Caspase-3）enzyme activity test results showed that the enzyme activity units of Hepa1-6 group and HepG2 group were（20.51±0.61）and（17.51±0.59）on the 7th day,which were statistically different from the control group of（22.15±1.14）.(P_Hepa1-6<0.05,P_HepG2<0.01),there was no statistically significant difference in enzyme activity between the Huh7 group（22.67±0.96）and the control group（22.15±1.14）（P>0.05）.Conclusion;The supernatant of hepatocellular carcinoma cells can improve the activity of the protoscoleces of Echinococcus granulosus in vitro,and the effect of HepG2 cell supernatant is more obvious.