二氧化碳作为膨宫介质在不同压力下对子宫内膜癌细胞增殖和凋亡影响的体外研究

Effect of CO_2 under different pressure as distending medium on proliferation and apoptosis of endometrial cancer cells in vitro

目的研究膨宫介质二氧化碳在不同作用压力下对子宫内膜癌细胞增殖和凋亡的影响。方法实验分为对照组和低(40 mmHg)、中(70 mmHg)、高(100 mmHg)压力3个实验组。实验组将人子宫内膜癌细胞(HEC-1-B)置于压力容器中,向容器内充入100%二氧化碳气体,使容器内压力分别达到40、70和100mmHg,各维持1 h。1 h后取出子宫内膜癌细胞,作好实验各组的压力标记,放入普通二氧化碳孵育箱继续培养。对照组则将子宫内膜癌细胞置于普通二氧化碳孵育箱环境中1 h。经上述处理后的连续7 d,每隔24 h采用四唑盐比色试验(MTT)测量各实验组和对照组子宫内膜癌细胞的增殖变化情况,采用流式细胞仪碘化丙啶(PI)活细胞染色法测量各实验组和对照组子宫内膜癌细胞凋亡变化的情况。结果①对照组、低压组、中压组、高压组内膜癌细胞MTT比色的OD值均呈逐渐上升趋势,对照组的上升趋势较实验各组明显,差异有显著性(P<0.05),低压组与中压组、低压组与高压组比较,差异有显著性(P<0.05),中压组与高压组的差异无显著性(P>0.05)。②对照组、低压组、中压组、高压组内膜癌细胞PI染色的凋亡细胞百分比均逐渐上升,实验各组的上升趋势较对照组明显,差异有显著性(P<0.05),低压组与高压组、中压组与高压组比较,差异有显著性(P<0.05),低压组与中压组比较,差异无显著性(P>0.05)。结论二氧化碳在40～100mmHg压力范围内,能够抑制子宫内膜癌细胞的增殖,促进子宫内膜癌细胞的凋亡,且随着压力的增加,二氧化碳对子宫内膜癌细胞增殖抑制和促进凋亡作用愈明显。

[Objective] To study the effect of distending medium CO2 on the proliferation and apoptosis of endometrial cancer ceils under different pressure. [Methods] Endometrial cancer cells were divided into low pressure （40 mmHg）, middle pressure （70 mmHg）, high pressure （100 mmHg）and control group. The cells in experimental groups were set into pressure containers. One hundred percent CO,_ gas was filled into the containers. The pressure in the containers of experimental groups was 40 mmHg, 70 mmHg or 100 mmHg which was kept for 1 hour, and then the cells were moved to normal CO2 incubators. The cells in the control group were set in the normal CO2 incubating container for 1 hour. After the above treatment, the optical density （OD） value of endometrial cancer cells was measured with MTT chromometry to explore the proliferation every 24 hour in the following 7 days. The apoptosis percentage of endometrial cancer cells was detected with PI staining flow cytometer method every 24 hour in the next 7 days. [Results] 1. The OD values in MTT of control group, low pressure group, middle pressure group and high pressure group all tended to rise. The tendency of the control group was more obvious than the experimental groups. There was statistical significance between control group and three experimental groups, between low and middle pressure groups, low and high pressure groups （P 〈0.05）;There was no statistical significance between middle and high pressure groups （P 〉0.05）. 2. In PI experiment, the apoptosis percentage of control, low pressure, middle pressure and high pressure groups gradually raised. The rising tendency of each experimental group was more obvious than that of the control group （P〈0.05）; There was also statistical significance between low and high pressure groups, middle and high pressure groups （P 〈0.05）.There was no statistical significance between low and middle pressure groups （P 〉0.05）. [Conclusion] CO2 can inhibit the proliferation and promote the apoptosis of the endometrial cancer cells with distending pressure from 40 mmHg to 100 mmHg. The higher the pressure is, the more obviously CO2 can inhibit the proliferation and promote the apoptosis of the endometrial cancer cells.