热CO_2气腹对胃癌细胞杀伤作用的实验研究

In vitro study of antitumor function of hyperthermic CO_2 pneumoperitoneum on human gastric cancer cells

目的:体外研究热CO2气腹对人胃癌细胞SGC-7901的杀伤作用,探讨临床上腹腔镜胃癌手术使用热CO2气腹抑制腹膜转移和戳孔种植的可行性。方法:建立体外热CO2气腹实验模型,人胃癌细胞SGC-7901经热CO2气腹(42℃、43℃,1～3 h)处理后,WST-8法检测细胞毒作用,流式细胞术定量分析、Hoechst 33342/PI荧光显微镜观察细胞形态,RT-PCR检测Bax、bcl-2基因mRNA的表达。结果:WST-8法检测显示热CO2气腹能显著抑制人胃癌细胞SGC-7901增殖,43℃作用3 h后,细胞存活率降至(65.83±2.40)%。流式细胞术及细胞形态学结果显示,诱导胃癌细胞凋亡是热CO2气腹作用的主要方式,与温度和作用时间呈正相关。RT-PCR检测结果显示,实验组(43℃,3 h)Bax基因mRNA表达水平显著高于对照组(25℃,3 h)差异有统计学意义(P<0.05);但对bcl-2的表达无明显影响。结论:热CO2气腹通过诱导细胞凋亡对人胃癌细胞有显著的增殖抑制作用,发生机制可能是通过上调Bax的表达促进细胞凋亡。

Objective： To in vitro observe the function of hyperthermic CO2pneumoperitoneum（HT-CO2）on human gastric cancer SGC-7901 cells,and then further evaluate the feasibility of inflating hyperthermic CO2 in gastric cancer patients when performing laparoscopic operation to inhibit peritoneal carcinomatosis and port site metastasis.Methods： Based on a newly desived in vitro experimental model,then the gastric cancer SGC-7901 cells were exposed to HT-CO2 at 42℃、43℃ for 1-3h.After dealing,the cytotoxicity was detected by WST-8 assay,and the death mechanism was assessed by Annexin V-FITC/PI flow cytometry,Hoechst 33342/PI fluorescent microscopy,and also the apoptosis-related gene mRNA expression of Bax and bcl-2 by real time fluorescent quantitative PCR.Results： Cytotoxicity detection showed that HT-CO 2 could significantly inhibit proliferation of gastric cancer cells（P0.05）,HT-CO 2（43 ℃,3 h）leaded to a decrease to（65.83±2.40）% in viable cells.Apoptosis detection and morphological analysis showed that HT-CO 2 could significantly induce apoptosis of gastric cancer cells in a temperature and time dependent way.Meanwhile,it was found that HT-CO 2（43 ℃,3 h）enhance the mRNA expression of Bax（P0.05）.However,the mRNA expression level of bcl-2 had no statistical difference by real time fluorescent quantitative PCR.Conclution： HT-CO 2 exerts a significant antitumor effect on gastric cancer cells through inducing apoptosis,and the mechanism involves the increase of the expression of Bax.