人工气腹对结肠癌细胞增殖影响的实验研究

The influence of artificial pneumoperitoneum on colon carcinoma cell proliferation in vivo and in vitro

目的研究不同气腹介质对人结肠癌细胞株LS-174T在体外及裸鼠体内增殖的影响，探讨腹腔镜结直肠癌手术的安全性。方法（1）细胞计数法绘制LS-174T细胞株生长曲线。（2）免疫组织化学检测细胞株HIF-1α、VEGF的表达。（3）丫啶橙／溴乙啶（AO／EB）双荧光染色检测细胞凋亡率。（4）流式细胞仪检测细胞周期的变化：（5）体内实验测量各组裸鼠实体肿瘤的重量及体积。结果体外试验中，经模拟CO2气腹作用后人结肠癌细胞LS-174T生长速度高于对照组，CO2、He人工气腹作用后结肠癌细胞中HIF-1α和VEGF的阳性表达率高于对照组，CO2组高于He组。He组细胞凋亡率高于CO2组和对照组。体内实验：CO2气腹组裸鼠移植瘤平均瘤重和瘤体积较免气腹组、He气腹组明显增高。结论裸鼠CO2气腹可促进结肠癌细胞LS-174T的生长，而He气腹促进肿瘤生长的能力较弱。

Objective To evaluate the safety of artificial pneumoperitoneum in laparoscopic colon carcinoma operation by an experimental study in which the influence of different pneumoperitoneal medium on colon carcinoma LS-174T cell proliferation was examined in vivo and in vitro in a nude mouse model. Methods 1. drawing cell growth curve by cytometry. 2. examing the expression of HIF-1α and VEGF by immunohistochemistry. 3. detecting cell apoptosis ratio by AO/EB double staining. 4. detecting cell life cycle by flow cytometry. 5. in vivo and in vitro measuring cell proliferation and the weight and volume of the implanted tumor tissue in each athymic mouse group. Results The in vitro growth speed and proliferating capacity of colon carcinoma cell line LS-174T under simulated CO2 pneumoperitoneum was higher than that in control group. The positive expression of HIF-1α and VEGF in colon carcinoma cells under CO2 and He artificial pneumoperitoneum was higher than that in control group, and the expression in CO2 group was higher than He group. Cell apoptosis ratio in He group was higher than control group. In the in vivo study, the weight and volume of implanted tumor in CO2 pneumoperitoneum group was significantly higher than without pneumoperitoneum group and He group. Conclusion CO2 pneumoperitoneum promotes the growth of colon carcinoma cell LS-174T, He pneumoperitoneum has relatively less significant effect on the growth of colon carcinoma cells.