体外模拟 CO2气腔对乳腺癌细胞 MMP-2和黏附分子 VCAM-1、ICAM-1表达的影响

Influence of artificial CO2 cavity on MMP-2 and VCAM-1, ICAM-1 expression in MDA-MB-231 cell

目的：探讨体外模拟CO2气腔对MDA-MB-231细胞基质金属蛋白酶-2（ matrix metalloprotei-nase 2，MMP-2）和黏附分子血管细胞间黏附分子-1（ vascular cell adhesion molecule 1，VCAM-1）、细胞间黏附分子-1（intercellular adhesion molecule 1，ICAM-1）表达的影响。方法体外建立人工气腔，MDA-MB-231细胞在7 mmHg（1 mmHg＝0.133 kPa）的CO2分压下暴露l、2及4 h，在处理后0、24、48及72 h，酶联免疫吸附法（ enzyme linked immunosorbent assay ， ELISA ）测定细胞培养液中 MMP-2浓度。流式细胞术测定VCAM-1、ICAM-1表达。缺氧组选用0 mmHg的氦气（1 h）。对照组为常规培养条件。结果1、2及4 h的CO2组处理后0 h时MMP-2的表达明显高于对照组（F＝15.045，P＜0.05）；2 h的CO2组处理后24 h时MMP-2的表达也明显高于对照组和1、4 h的CO2组（F＝5.976，P＜0.05）。1、2及4 h的CO2组处理后0、24 h时VCAM-1的表达显著高于对照组（F1＝18.321，F2＝20.443，P＜0.05）；4 h的CO2组处理后72 h时VCAM-1的表达显著低于1、2 h的CO2组（F＝15.045，P＜0.05）。缺氧组和1、2及4 h的CO2组处理后0 h时ICAM-1的表达显著高于对照组，其中2 h的CO2组表达又高于1、4 h的CO2组（F＝73.765，P＜0.05）；2、4 h的CO2组处理后24 h时ICAM-1的表达显著高于对照组和1 h的CO2组（F＝46.322，P＜0.05）；2 h的CO2组处理后48 h时ICAM-1的表达显著高于对照组和1、4 h的CO2组（F＝22.315，P＜0.05）。结论模拟7 mmHg的CO2气腔可使MDA-MB-231细胞MMP-2、VCAM-1、ICAM-1的表达增高，乳腔镜CO2气腔可能对乳腺癌细胞的转移具有一定影响力。

Objective To investigate the influence of in vitro artificial CO 2 cavity on matrix metallopro-teinase 2（MMP-2）, adhesion molecule vascular cell adhesion molecule 1（VCAM-1）, and intercellular adhesion molecule 1（ICAM-1）expression in MDA-MB-231 cell.Methods An in vitro artificial CO2 cavity model was es-tablished.MDA-MB-231 cells were exposed to CO2 under the pressure of 7 mmHg for 1, 2 and 4 hours, respective-ly.MMP-2 concentration was measured by enzyme linked immunosorbent assay （ELISA）.VCAM-1 and ICAM-1 ex-pression were measured by flow cytometry 0, 24, 48 and 72 hours after CO2-insufflation.Hypoxia group was ex-posed to 0 mmHg helium for 1 h, and the control group was exposed to 37℃incubator only .Results Compared with that in the control group , MMP-2 expression in the 1, 2 and 4 hours treatment group was significantly elevat-ed at 0 hour（F=15.045, P〈0.05）, and the MMP-2 expression in the 2 hours CO2 treatment group was signifi-cantly elevated after 24 hours compared with that in the control group and 1, and 4 hours CO2 treatment group （F=5.976, P〈0.05）.The VCAM-1 expression was significantly elevated at 0 hour and after 24 hours in the 1, 2 and 4 hours CO2 treatment group compared with that in the control group （ F1 =18.321, F2 =20.443, P〈0.05）, and significantly declined after 72 hours in the 4 hours CO2 treatment group compared with that in 1, and 2 hours CO2 treatment group（F=15.045,P〈0.05）.ICAM-1 expression was significantly elevated at 0 hour in hypoxia group and 1, 2, 4 hours CO2 treatment group compared with that in the control group , Meanwhile it was higher in 2 hours CO2 treatment group than in 1 hours and 4 hours CO2 treatment group（F=73.765, P〈0.05）. ICAM-1 expression was significantly elevated after 24 hours in 2 and 4 hours CO2 treatment group compared with that in the control group and 1 hour CO2 treatment group（F=46.322, P〈0.05）, and it was significantly elevat-ed after 48 hours in 2 hours CO2 treatment group compared with that in the control group and 1, and 4 hours CO2 treatment group（F=22.315, P〈0.05）.Conclusion The expression of MMP-2, adhesion molecule VCAM-1, and ICAM-1 in MDA-MB-231 cells is elevated after exposure to artificial 7 mmHg CO2 cavity, and CO2 cavity of mastoscopy may modulate the metastasis capacity of breast tumor cells .