不同压力CO_2气腹对胃癌细胞增殖凋亡的影响

Effects of different pressures of CO_2 pneumoperitoneum on proliferation and apoptosis of gastric cancer cells

目的通过体外模拟气腹环境,观察CO2在不同压力下对胃癌细胞MKN-45增殖、凋亡的影响。方法以CO2为气体介质,在气腹机作用下维持密闭培养箱内压力为0、5、10、15mmHg,作用时间为4h。于处理后用血气分析仪检测培养液pH值;MTT法检测细胞增殖情况;通过流式细胞仪观察细胞凋亡指数变化;采用AnnexinV-FITC/PI双标染色、流式细胞仪测定凋亡细胞比例。结果处理结束后即刻检测,CO2组细胞培养液呈酸性,但在恢复正常培养后6h即恢复正常。MTT法检测细胞增殖变化,CO2组0、5、10mmHg压力下处理组与对照组比较无明显差异(P>0.05),而在15mmHg压力下处理组与对照组比较OD490值在前3d无明显差异(P>0.05),在4～7dOD490值下降非常显著(P<0.01)。在CO2环境下,0mmHg组细胞凋亡指数和凋亡比例与对照组相比无明显差异(P>0.05),而5、10、15mmHg组明显大于正常对照组(P<0.05)。结论在较低压力范围内(≤10mmHg)CO2对胃癌细胞增殖、凋亡影响不大,而在15mmHg压力下,CO2可抑制MKN-45细胞增殖,促进其凋亡。

Objective This study aimed to investigate the influence of CO2 pneumoperitoneum administered at different pressures on the proliferation and apoptosis of human gastric cancer cells in vitro. MethodsGastric cancer cells MKN-45 were cultured in the CO2 environment at different pressures （0, 5, 10, 15 mmHg） in vitro. After 4 hours, pH of the culture media were measured respectively in each group.The proliferation of MKN-45 cells was examined by MTT assay respectively in each group. Apoptosis of MKN-45 cells was investigated using annexin Ⅴ-FITC/PI double labeled staining and tested by flow cytometry. Results pH was recovered to normal after 6 hours in the CO2 group. There was no significant difference between the CO2 group （ under 10 mm Hg ） and the control group（P0.05） on the proliferation of MKN-45 cells . The cultured cells exposed to 15 mm Hg in the CO2 environment grew slower than in the control group at 4 to 7 days （P0.01=but there was no significant difference at 1 to 3 days（P0.05）. There was no significance in apoptotic index of MKN-45 cells between the 0 mm Hg CO2 group and control group（P0.05）, and apoptotic indexes of MKN-45 cells in 5, 10, 15 mm Hg CO2 groups were upregulated than those in the control group（P0.05）. Conclusion There were no obvious effects on the proliferation of gastric cancer cells under 10 mm Hg CO2 insufflation pressure. Inhibited proliferation and enhanced apoptosis of human gastric cancer cells were achieved under 15 mm Hg CO2 insufflation pressure.