CO2气腹对胃癌细胞周期及细胞周期蛋白的影响

目的 通过检测不同压力CO2气腹环境下胃癌细胞周期及其相关蛋白表达的变化,探讨CO2气腹对胃癌细胞生长增殖的影响.方法 将胃癌MKN-45细胞置于0、10、12和15 mm Hg CO2气腹环境下培养4 h,然后用流式细胞法检测胃癌细胞周期的变化,再用Western blot方法检测细胞周期蛋白Cyclin D1、CDK4、Rb和pRb的表达,最后用免疫沉淀法检测细胞Cyclin D1/CDK4结合力.结果 0、10、12 mm Hg CO2气腹组胃癌细胞增殖指数与对照组比较差异均无统计学意义(P>0.05),15 mm Hg CO2气腹组胃癌细胞增殖指数(27.4%4±3.7%)与对照组(36.4%±3.3%)比较显著下降(P<0.05).0、10、12 mm Hg CO2气腹组CDK4、Cyclin D1、pRb蛋白表达以及Cyclin D1和CDK4的结合能力与对照组比较差异均无统计学意义(P>0.05),15 mm Hg CO2气腹组CDK4、Cyclin D1、pRb蛋白表达以及Cyclin D1和CDK4的结合能力(0.71%±0.12%、0.93%±0.21%、0.54%±0.11%、0.18%±0.02%)与对照组(1.05%±0.16%、1.40%±0.24%、0.75%±0.14%、0.31%±0.02%)比较显著下降(P<0.05).0、10、12、15 mm Hg CO2气腹组Rb蛋白表达与对照组比较差异无统计学意义(P>0.05).结论 临床常用压力的CO2气腹对胃癌细胞周期无显著影响,15 mm Hg CO2气腹抑制细胞增殖,其原因可能与Cyclin D1和CDK4的表达下调有关.

Effects of CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line

Objective To investigate the effects of different CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line. Methods Human gastric cancer cell line MNK-45 were exposed to 0,10,12 and 15 mm Hg CO2 pneumoperitoneum in vitro for 4 hrs. Cell cycle was measured by flowcytometry, the expression of CDK4 ,Cyclin D1、Rb and pRb was studied by Western-blot, and the binding ability of CDK4 and Cyclin D1 was evaluated by immunoprecipitation. Results The cell proliferation index in 15 mm Hg CO2 pneumoperitoneum group dropped significantly (27.4% ± 3. 7%) vs. (36. 4% ±3. 3%) ,P <0. 05, while that in other groups did not change significantly. The protein of CDK4、Cyclin D1and binding ability of Cyclin D1 and CDK4 dropped dramatically in the 15 mm Hg CO2 pneumoperitoneum group (0.71%±0.12%),(0.93% ±0.21%),(0.54%±0.11%),(0.18% ±0.02%) vs. (1.05% ±0.16%),(1.40% ±0.24%),(0.75% ±0.14%),(0.31% ±0.02%), all P<0.05. There were no changes of Rb in protein levels, while the phosphorylated Rb dropped obviously. Conclusion There was no obvious effects of clinical CO2 pneumoperitoneum on gastric cancer cells growth and proliferation.