不同压力的二氧化碳气腹对胃癌细胞迁移运动和细胞骨架的影响

Effects of different CO2 pressure pneumoperitoneum on the migration and cytoskeleton in gastric cancer cells

目的通过体外模拟气腹环境，观察不同压力CO2或He（helium）对胃癌细胞MKN-45迁移运动和细胞骨架的影响。方法将MKN-45细胞置于充满CO2或He的密闭培养箱中，按模拟气腹种类不同分为对照组、CO2气腹组和He气腹组，模拟气腹压力分别为12mmHg和15mmHg，作用时间均为4h。于处理结束后即刻用血气分析仪检测培养液pH值；Transwell法观察细胞迁移运动变化；激光共聚焦显微镜观察细胞骨架变化。结果CO2组细胞培养液呈酸性，He组细胞培养液呈碱性。CO2组和He组在12mmHg压力下MKN-45细胞穿过滤膜的数量较对照组差异无统计学意义（P〉0．05）。CO2 15mmHg组细胞穿过滤膜的数量较对照组明显减少（P〈0．01）；He15mmHg组与对照组差异无统计学意义（P〉0．05）；CO2 15mmHg组细胞微丝、微管结构模糊，伪足消失。结论在临床常用气腹压力（12mmHg）下CO2对胃癌细胞迁移运动及细胞骨架无明显影响；在15mmHg压力下，CO2对胃癌细胞迁移运动起抑制作用，其作用机制可能与其细胞骨架受破坏有关。

Objective To investigate the influence of CO2 and He insufflation administered at different pressures on the migration and cytoskeleton of cultured human gastric cancer cells. Methods The cultured gastric cancer cells MKN-45 were exposed to a CO2 or He environment maintained at different pressures （12, 15 mm Hg）. After 0, 2, 4, 6, 8 hours exposure to CO2 or He environment, pH of the MKN-45 cells culture media was measured with blood gas analysis. The cell migration was detected with Transwell technology. The cell cytoskeleton was observed with laser confocal microscope. Results The media pH was acid after exposure to CO2 environment, while it was basic in the He group. The number of cells passing millipore in 12 mm Hg CO2 or He insufflation pressure were not significantly different with control group （P〉0.05）, however in 15 mm Hg pressure CO2 group, it was significantly decreased as compared to control group （P〈0.01）. The microfilament and microtubule in gastric cancer cell were ambiguous in 15 mm Hg pressure CO2 group. Conclusions There are no obvious effects on the migration and cytoskeleton of MKN-45 cells under 12 mm Hg CO2 insufflation pressure. The migration and cytoskeleton of MKN-45 cells can be inhibited in 15 mm Hg CO2 pneumoperitoneum environment.