泡状棘球蚴原头节与肝癌细胞共培养的相互作用研究

目的 探究泡状棘球蚴原头节与肝癌细胞共培养后,原头节的活性、成囊以及肝癌细胞的增殖与凋亡情况.方法 构建泡状棘球蚴原头节与肝癌细胞共培养模型,以存在肝癌细胞共培养的泡球蚴原头节为实验组,无肝癌细胞共培养的泡球蚴原头节为对照组,光镜下观察泡状棘球蚴原头节存活数量、活性以及成囊情况;以存在原头节共培养的肝癌细胞为实验组,无...

Study of the interaction between co-cultured alveolar echinococcosis protoscolex and hepatoma cells

To investigate the viability and cyst formation of alveolar echinococcosis protoscolex co-cultured with hepatoma cells,and the proliferation and apoptosis of hepatoma cells,we established a co-culture model.Alveolar echinococcosis protoscolex co-cultured with hepatoma cells was the experimental group,and alveolar echinococcosis protoscolex co-cultured without hepatoma cells was used as a control.The number of surviving cells,viability and cyst formation of alveolar echinococcosis protoscolex were observed under a light microscope.After co-culture,the survival rate of alveolar echinococcosis protoscolex in the experimental group(91.42±2.47)%]was significantly higher than that in the control group(73.10±4.46)%,t=-7.738,P<0.05].When hepatoma cells were used as feeder cells,the cyst formation rate of the protoscolex was(66.72±2.02)%,a rate higher than that of non-hepatoma cells(30.21±2.25)%,t=11.228,P<0.01].The hepatoma cells co-cultured in protoscolex were used as the experimental group,and the hepatoma cells without protoscolex were used as the control group.The proliferation of hepatoma cells was detected with CCK-8 assays,which showed that,with an increasing proportion of protoscolex,the proliferation ability of hepatoma cells was gradually enhanced and was significantly higher than that of control cells.Apoptosis was detected with Annexin V-FITC/PI flow cytometry,and the mRNA expression of the apoptosis-associated molecules caspase3,caspase9,Bax,Bcl-2,P21 and P53 was detected by real-time quantitative PCR.The expression of Bcl-2 in the experimental group was significantly higher than that in the control group,whereas the expression of caspase3,caspase9,Bax,P21 and P53 was lower than that in the control group.The protein expression of caspase3,caspase9,Bax and Bcl-2 was detected by western blotting.The results were consistent with those of real-time quantitative PCR,which showed that the expression of Bcl-2 in the experimental group was significantly higher than that in the control group,whereas the expression of caspase3,caspase9 and Bax in the experimental group was lower than that in the control group.Thus,when alveolar echinococcosis protoscolex is co-cultured with hepatoma cells,the hepatoma cells promote the survival of the protoscolex,maintain the high viability of the protoscolex and promote cyst formation;the protoscolex also promotes the proliferation of hepatoma cells and inhibits apoptosis in liver cancer cells.