不同压力二氧化碳气腹对胃癌细胞增殖运动的影响

Effects of CO2 pneumoperitoneum with different pressures on proliferation and motility of gastric cancer cells MKN-45

目的 通过体外模拟气腹环境，观察不同压力CO2对胃癌细胞MKN-45增殖、运动的影响。方法 将MKN-45细胞置于充满CO2的密闭培养箱中，按CO2压力不同分为对照组、0、5、10和15mm Hg组（1mm Hg=0．133 kPa），作用时间均为4h。用血气分析仪检测培养液pH值；MTT法检测细胞增殖情况；流式细胞仪观察细胞周期变化；Transwell法观察细胞迁移运动变化。结果 0、5、10和15mm Hg组细胞培养液在0h呈酸性。MTT法检测细胞增殖发现0、5、10mm Hg组与对照组比较差异无统计学意义（P〉0．05），15mm Hg组与对照组比较吸光度（OD）值在1—3d差异无统计学意义（t=0．625，-0．537，0．137，P〉0．05），在4—7d差异有统计学意义（t=26．622，13．376，18．839，7．595，P〈0．01）。0、5、10mmHg组细胞增殖指数和细胞穿过滤膜的数量与对照组比较差异无统计学意义（t=1．134，-0．538，-0．829；t=0．330，0．794，0．508，P〉0．05），15mmHg组与对照组比较差异有统计学意义（t=11．225，8．775，P〈0．01）。结论 在15mmHg压力下，CO2对MKN-45细胞增殖、运动起抑制作用。

Objective To investigate the effects of CO2 pneumoperitoneum with different pressures on the proliferation and motility of cultured human gastric cancer cell line MKN-45 in vitro. Methods According to different CO2 pressures, gastric cancer cell line MKN-45 was divided into 0 mm Hg group, 5 mm Hg group, 10 mm Hg group, 15 mm Hg group and control group. The pH value of cell culture medium, proliferation, changes of cycle and migration of gastric cancer cells was detected by blood gas analyzer, MTT assay, flow cytometry and Transwell method, respectively 4 hours later after CO2 insufflation. Results The pH values of culture medium in 0 mm Hg, 5 mm Hg , 10 mm Hg and 15 mm Hg group were lower than 7 at hour 0. No statistical difference of gastric cancer cell proliferation was seen between 0 mm Hg, 5 mm Hg, 10 mm Hg group and control group （P 〉 0.05）. The difference of optical density had no statistical difference between 15 mm Hg group and control group from day 1 to day 3 （ t = 0.625, - 0.537, 0.137, P 〉 0.05）, but had statistical difference from day 4 to day 7 （ t = 26. 622, 13. 376, 18. 839, 7. 595, P 〈0.01 ）. In regard of cell proliferation and number of cells migrated through the filter membrane, there was no statistical difference between groups 0 mm Hg, 5 mm Hg, 10 mm Hg and control group （ t = 1.134, - 0.538, - 0. 829 ; t = 0. 330, 0.794, 0. 508, P 〉 0.05）, but significant statistical difference between 15 mm Hg group and control group （ t = 11. 225, 8. 775, P 〈0.01）. Conclusions CO2 pneumoperitoneum has inhibitory effect on proliferation and motility of gastric cancer cell line MKN-45 under 15 mm Hg pressure.