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℃,1～3h)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3h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℃,3h)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.%Objectices: To in vitro observe the function of hyperthermic CO2 pneumoperitoneum (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 ex perimental 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 micrpscopy,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-CO2 could significantly inhibit proliferation of gastric cancer cells(P<0.05), HT-CO2(43℃,3 h) leaded to a decrease to(65.83 ± 2.40)% in viable cells. Apoptosis detection and morphological analysis showed that HT-CO2 could significantly induce apoptosis of gastric cancer cells in a temperature and time dependent way. Meanwhile,it was found that HT-CO2(43℃,3 h)enhance the mRNA expression of Bax(P<0.05). However, the mRNA expression level of bcl-2 had no statistical difference by real time fluorescent quantitative PCR. Conclution: HT-CO2 exerts a significant antitumor effect on gastric cancer cells through inducing apoptosis, and the mechanism involves the increase of the expression of Bax.
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