CO2气腹对胃癌MKN-45细胞黏着斑激酶的影响

Effects of CO2 pneumoperitoneum on focal adhesion kinase of gastric cancer MKN-45 cells

目的研究CO2气腹对胃癌MKN-45细胞黏着斑激酶（FAK）的影响。方法体外模拟不同压力CO2气腹环境，实验组：人胃癌MKN-45细胞置于5、10、15mmHg（1mmHg=0．133kPa）CO2气腹环境培养4h。对照组：常规条件培养人胃癌MKN-45细胞。采用Western blot法检测各组细胞FAK及磷酸化FAK（FAK Tyr397）的表达量，且分别观察15mmHg CO2作用0．5、2、4h的情况。多组比较采用单因素方差分析，组间比较采用LSD法检验。结果实验组5、10、15mmHg CO2气腹环境下，FAK表达量分别为2．14±0．17、2．07±0．21、2．52±0．26；FAK Tyr397表达量分别为1．82±0．28、1．93±0．52、3．71±0．37；而对照组FAK表达量为2．43±0．46，FAK Tyr397表达量为1．71±0．23，两组比较差异有统计学意义（F=2．171，26．951，P〈0．01）。15mmHg CO2作用0．5、2、4h后，FAKTyr397表达量分别为3．41±0．44、4．12±0．56、5．24±0．41，三者比较差异有统计学意义（F=116．119，P〈0．01）。脱离气腹后恢复至处理前水平（0．72±0．16）。结论不同压力CO2气腹环境处理胃癌MKN-45细胞4h不增加其FAK表达，但可使其磷酸化而激活，且压力越高，作用时间越长，磷酸化程度越高，但脱离气腹环境后，其活性迅速降至处理前水平。

Objective To investigate the effects of CO2 pneumoperitoneum on the expression of focal adhesion kinase （FAK） of gastric cancer MKN-45 cells. Methods CO2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg （ 1 mm Hg =0. 133 kPa） ] for 4 hours. The condition of the cells exposed to CO2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK （ FAK Tyr397 ） of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14 ± 0.17, 2.07 ± 0.21 and 2.52 ± 0.26, respectively, and the expression of FAK Tyr397 was 1.82 ± 0.28, 1.93 ± 0.52 and 3.71 ± 0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43 ± 0.46 and 1.71 ± 0.23, respectively. We found significant differences between the 2 groups （ F = 2. 171, 26. 951, P 〈 0.01 ）. After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41 ±0.44, 4.12 ± 0.56 and 5.24 ± 0.41 respectively, which is also significantly different （ F = 116. 119, P 〈0.01 ）. The expression of FAK Tyr397 was back to 0.72 ± 0.16 1 hour after the release of CO2. Conclusions CO2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.