热CO2处理后树突状细胞源性外体对胃癌AGS细胞增殖的抑制作用

Inhibitory effect of Dexosome from dendritic cells treated with heat-CO2 pneumoperitoneum of pro- liferation of human gastric adenocarcinoma AGS cells

目的观察热CO2处理后树突状细胞（DC）来源外体（Dex）对胃癌AGS细胞增殖的抑制作用，探讨腹腔镜胃癌根治术时引入热CO2处理的作用效果。方法建立热CO2气腹体外实验模型，分组处理DC后，提取Dex并通过透射电镜和Westernblot鉴定。将各组Dex与AGS细胞共培养后，以细胞计数试剂盒（CCK-8）法检测AGS细胞增殖，Westernblot检测Dex中热休克蛋白70（HSP70）的表达。结果透射电镜显示Dex为直径30～100nm的膜性微囊，其CD63蛋白表达明显高于全细胞裂解物（P〈0．05）。单纯热处理组与热CO2气腹组Dex对AGS细胞增殖抑制率分别为（20．50±3．07）％和（45．30±2．13）％，两组间及两组与对照组问比较差异有统计学意义（P〈0．05）。与对照组比较，热CO2气腹处理组Dex中HSP70表达明显增高（P〈0．05）。结论成功提取Dex，热CO2处理后Dex能显著抑制胃癌AGS细胞增殖，可能与Dex中HSP70表达增高有关。

Objective To observe the inhibitory effects of dendritic cells （DCs）-derived Dexo- some （Dex） on proliferation of human gastric adenocarcinoma cell line AGS after treated with heat-CO2 and investigate the value of heat-CO2 pneumoperitoneum used during laparoscopic surgery. Methods An in vitro heat-CO2 pneumoperitoneum experimental box was built. After heat-CO2 pneumoperitoneum treatment, the DCs-derived Dex was isolated and identified through transmission electronic microscope （TEM） and Western blotting. AGS cells were co-cultured with Dex, and the cell proliferation was measured by cell counting kit-8 （CCK-8）. The expression of heat shock protein 70 （HSPT0） in Dex was examined by West- ern blotting. Results The Dex from DCs culture supernatants was revealed to be 30 to 100 nm membrane vesicles by TEM, and its expression of marker CD63 was significantly higher than whole cell lysates （P 〈 0. 05 ）. The proliferation inhibition rate of AGS cells was （ 20. 50 ± 3.07 ） % in heat treated group and （45.30±2. 13）% in heat-CO2 treated group respectively （P 〈 0. 05）. The expression of HSPT0 in Dex treated with heat-CO2 was significantly higher （ P 〈 0. 05 ）. Conclusion Dex was isolated successfully. Proliferation of AGS cells could be inhibited significantly by Dex after DCs treated through Heat-CO2, which may be related to the over-expression of HSPT0 molecules in Dex.