七叶皂苷钠对肝癌细胞迁移黏附能力的影响

Effect of sodium aescinate on migration and adhesion ability of hepatocarci-noma cells

目的:研究七叶皂苷钠对HepG2肝癌细胞迁移和黏附能力的影响。方法:体外培养HepG2细胞,低、中、高浓度实验组细胞分别给予20、40、60μmol/L七叶皂苷钠,采用细胞缓慢聚集实验观察七叶皂苷钠对悬浮细胞缓慢能力的影响,采用细胞分离试验观察七叶皂苷钠对细胞分离能力的影响,采用细胞划痕实验观察七叶皂苷钠对细胞迁移能力的影响,采用Western blot方法观察七叶皂苷钠对HepG2细胞内E-钙黏蛋白(E-cadherin)表达的影响。结果:对照组细胞聚集团块体积小,数量多且疏松,随着七叶皂苷钠浓度的升高,细胞团块的体积有所增大,数量减少但致密度增加。低浓度实验组细胞团数为24.19±2.94,中浓度实验组细胞团数为13.93±3.18,高浓度实验组细胞团数为2.14±1.01,均低于对照组的细胞团数(64.28±6.83,P<0.05),且高浓度实验组细胞团数低于中浓度实验组(P<0.05),中浓度实验组的细胞团数低于低浓度实验组的细胞团数(P<0.05)。相对于对照组,随着七叶皂苷钠浓度的升高,HepG2的细胞聚集能力升高,应用1 mmol/L CaCl2的胰酶处理细胞后,形成团块体积较大(P<0.05),且团块体积大小与药物含量成正比。细胞间粘附能力随药物浓度增加而加强,随药物浓度升高,NTC/NTE降低(P<0.05),低浓度实验组NTC/NTE为0.50±0.06,中浓度实验组NTC/NTE为0.35±0.03,高浓度实验组NTC/NTE为0.16±0.04,均小于对照组NTC/NTE(0.90±0.09,P<0.05)。随着药物浓度升高,HepG2细胞迁移能力受限,各浓度实验组细胞划痕宽度均宽于对照组,且药物浓度越高,划痕宽度越大(P<0.05)。对照组细胞E-cadherin蛋白表达低于各浓度实验组,且随着药物浓度升高,E-cadherin蛋白表达增高(P<0.05)。结论:细胞实验显示七叶皂苷钠能够增强HepG2细胞的黏附能力,降低细胞迁移能力,能够降低肝癌扩散转移风险,且随着七叶皂苷钠剂量的升高,其对HepG2肝癌细胞的作用效果逐步增强。

Objective:To study the effect of sodium aescinate on the migration and adhesion of HepG2 liver cancer cells.Methods:HepG2 cells were cultured in vitro and treated with 20, 40 and 60 μmol/L sodium aescinate in the low, medium and high concentration groups respectively.The slow cell aggregation experiment was used to characterize the effect of sodium aescinate on the slow aggregation ability of suspension cells.While the cell separation experiment was used to study the effect of sodium aescinate on the cell separation ability;and application Scratch experiment was used to characterize the effect of sodium aescinate on cell migration ability.Western blot experiment was used to characterize the effect of sodium aescinate on the expression of E-cadherin in liver cancer cells.Results:With the increase in the concentration of sodium aescinate, the volume of cell clumps increased, the number decreased but the density increased.However in the control group, the cell clumps were small, large in number and loose.After counting and analysis, the number of cell clusters in the control group was 64.28±6.83, the number of cell clusters in the low concentration experimental group(20 μmol/L aescinate sodium) was 24.19±2.94,and the medium concentration experimental group(40 μmol/L aescin) The number of cell clusters in sodium) was 13.93±3.18,and the number of cell clusters in the high-concentration experimental group(20 μmol/L aescinate sodium) was 2.14±1.01,which were lower than those in the control group(P<0.05).And the number of cell clusters in the high concentration experimental group was lower than that in the medium concentration experimental group(P<0.05),and the number of cell clusters in the medium concentration experimental group was lower than that in the low concentration experimental group(P<0.05).The results showed that with the increase of the concentration of sodium aescinate,the cell aggregation ability of Hep G2 increased,and it was higher than that of the control group(P<0.05).After treatment with 1 mmol/L CaCl2 trypsin,the mass size was larger(P<0.05),and the mass size was proportional to the drug content.After the cells were treated with 1 mmol/L EDTA trypsin,the cells of each group were dispersed into single cells,and the dispersion degree of the cells of each group was similar(P>0.05).The intercellular adhesion ability was enhanced with the increase of drug concentration,and the NTC/NTE ratio was decreased with the increase of drug concentration,the NTC/NTE of the low concentration experimental group was 0.50±0.06,the NTC/NTE of the medium concentration experimental group was 0.35±0.03,the NTC/NTE of the high concentration experimental group was 0.16±0.04,and the NTC of the control group/NTE was 0.90±0.09.The NTC/NTE of the drug group was lower than that of the control group(P<0.05).With the increase of drug concentration,the migration ability of HepG2 cells was limited.The scratch width of cells in each concentration experimental group was wider than that in the control group,and the higher the drug concentration,the larger the scratch width(P<0.05).The expression of E-cadherin protein in the control group was lower than that in the experimental groups,and with the increase of drug concentration,the expression of E-cadherin protein increased(P<0.05).Conclusion: Sodium aescinate can enhance the adhesion ability of HepG2 liver cancer cells,reduce the migration of liver cancer cells,and reduce the risk of spreading and metastasis of liver cancer.As the dose of sodium aescinate increases,its effect on HepG2 liver cancer cells is more effective.