模拟气腹对肝癌SMMC-7721细胞侵袭的影响

Effects of CO_2 Pneumoperitoneum on Invasion of Hepatoma Carcinoma Cells Line SMMC-7721

目的:通过体外模拟气腹环境,观察不同压力CO2对肝癌SMMC-7721细胞侵袭能力的影响。方法:建立体外气腹模型,将人肝癌细胞株SMMC-7721分别在5mmHg、10mmHg、15mmHg不同压强CO2气腹环境及常规培养环境中暴露2h,然后继续常规培养。用Transwell侵袭小室检测细胞体外侵袭能力的变化;黏附试剂盒及免疫细胞化学方法检测不同时间点细胞黏附能力及基质金属蛋白酶9(MMP-9)、血管内皮生长因子(VEGF)蛋白表达。结果:肝癌SMMC-7721细胞侵袭、黏附能力及MMP-9蛋白表达在15mmHg组中比在对照组中显著提高(P<0.05);VEGF蛋白表达在10mmHg组和15mmHg组中比在对照组中均显著提高(P<0.05);细胞黏附表达及MMP-9、VEGF蛋白表达在CO2气腹作用后逐渐升高,36h达高峰,随后下降。结论:较高压力CO2气腹环境对肝癌SMMC-7721细胞的侵袭能力有显著促进作用,可能与CO2气腹显著促进肝癌SMMC-7721细胞MMP-9及VEGF蛋白表达有关。

Objective：To investigate the effects of carbon dioxide（CO2） pneumoperitoneum environment with different pressures on invasion ability of cultured human hepatoma carcinoma cells line SMMC-7721 in vitro.Methods： With an in vitro artificial pneumoperitoneum model,human hepatoma carcinoma cells line SMMC-7721 were exposed to CO2-insufflation of three pressure groups（5,10 and 15 mmHg） and the control group.After exposion,transwell chamber was used to determine the invasion capacity of hepatoma carcinoma cells in vitro.The different time point after exposion,adhesive kit and immunocytochemical stain was respectly used to detected the adhesive ability and the expressions of MMP-9 and VEGF of human hepatoma carcinoma cells.Results： The invasion,adhesive ability and The MMP-9 expressions of hepatoma carcinoma cells line SMMC-7721 in 15 mmHg group showed significantly enhanced than in control group（P0.05）.The VEGF expressions in 15 mmHg and 10mmHg groups showed significantly raised than in control group（P0.05）.The adhesive ability of cells and the protein expressions of MMP-9 and VEGF rised gradually and reached a peak at 36 h after exposition,then decreased.Conclusions： High pressure CO2 pneumoperitoneum envioronment can significantly promote the invasion ability of hepatoma carcinoma cells line SMMC-7721.The mechanism may due to obviously promotion of the expression of MMP-9 and VEGF.