体外模拟CO_2气腹对人结肠癌SW480细胞生长的影响

Effects of Simulated CO_2 Pneumoperitoneum on the Growth of Human Colonic Cancer SW480 Cells

目的通过建立体外模拟CO2气腹环境,研究CO2气腹对人结肠癌SW480细胞生长的影响。方法建立体外模拟CO2气腹环境,在全自动气腹机作用下维持密闭培养箱内压力分别为9、12、15 mm Hg,持续时间分别为1、2、4 h。在处理后第12 h通过流式细胞仪观察细胞周期变化;在处理后第12、24、36、48、60、72 h用MTT法检测细胞增殖情况;在处理后0、0.5、1、1.5、2、2.5 h通过电子pH计检测细胞培养液pH值的变化。结果流式细胞仪检测细胞周期结果显示,CO2气腹组G0/G1期细胞比例与对照组差异无统计学意义(P>0.05)。CO2气腹压力对于人结肠癌G0/G1期细胞周期有显著影响(P=0.008),但对S期、G2/M期无影响(P>0.05);CO2气腹持续作用时间对各细胞周期无影响(P>0.05)。MTT法检测细胞增殖结果显示,各气腹组人结肠癌SW480细胞的D(490)值均低于对照组,且9 mm Hg-4 h、12 mm Hg-4 h、15 mm Hg-2 h气腹组在处理后第24、36 h,15 mm Hg-4 h气腹组在处理后第24、36、48、60 h的人结肠癌SW480细胞的D(490)值与对照组差异具有统计学意义(P<0.05)。压力处理后第24、36 h对SW480细胞的增殖有显著的一过性影响(P<0.01),而在第12、48、60、72 h对SW480细胞的增殖无影响(P>0.05);作用时间对SW480细胞的增殖无影响(P>0.05)。电子pH计检测细胞培养液pH值结果显示,除9 mm Hg-1 h气腹组在处理后2.5 h细胞培养液的pH值与对照组无统计学差异外(P>0.05),其余各气腹组在处理后0、0.5、1、1.5、2、2.5 h与对照组差异有统计学意义(P<0.05)。结论体外模拟CO2气腹环境,不会促进结肠癌细胞的增殖,且高CO2气腹压力对结肠癌细胞的增殖有一过性抑制作用,CO2气腹持续时间对结肠癌细胞的增殖无影响。

Objective To study the effects of simulated CO2 pneumoperitoneum on the growth of cultured human colonic cancer SW480 cells. Methods Colonic cancer SW480 cells were exposed to CO2 environment for different time durations （ 1, 2 and 4 hours） under different pressures （9, 12 and 15 mm Hg）, and the cells that cultured in normal experiment were set as a control group. The cell cycle were examined by flow cytometry after being moved to normal conditions for 12 h; and the proliferation of SW480 cells were examined by MTT after being moved to normal conditions for 12, 24, 36, 48, 60 and 72 h respectively; and pH of the culture media was measured after being in normal condition for 0, 0.5, 1, 1.5, 2 and 2.5 h. Results The G0/G1 ratio showed no statistical difference between CO2 pneumoperitoneum and control groups （P 〉 0.05 ）. CO2 pneumoperitoneum pressure had significant influence on the numerical values of human colonic cancer cells in G0/G1 phase （P = 0. 008） , however no influence on the numerical values of the cells in S and G2/M phases（P 〉 0.05）. The duration of exposure to simulated CO2 pneumoperitoneum had no significant influence on the cell cycle （P 〉 O. 05）. MTT showed that the proliferative viability in CO2 group grew slower than that in the control; the 9 mm Hg-4 h, 12 mm Hg-4 h, 15 mm Hg-2 h CO2 groups at hours 24 and 36, and 15 mm Hg-4 H CO2 group at hours 24, 36, 48 and 60 hours showed significantly different D（490） values compared to the control （ all P 〈 0.05 ）. CO2 pneumoperitoneum pressure had transient effect on the proliferation of the SW480 ceils at hours 24 and 36 （P 〈 0.01 ）, but showed no influence at hours 12, 48, 60, and 72 （ P 〉 0.05 ）. All the experiment groups, except for group 9 mm Hg-1 h （ P 〉 0.05 ）, showed significantly higher pH values at hours 0.5, 1, 1.5, 2, and 2.5 （ all P 〈 0.05 ） compared with control groap. Conclusions Simulated CO2 pneumoperitoneum does not promote the growth of colonic cancer SW480 cells. The insufflation pressure restrains the growth of colonic cancer SW480 cells for a short period, but the duration of CO2 pneumoperitoneum does not affect the growth of colonic cancer SW480 cells.