热CO2处理树突状细胞源性外泌体诱导胃癌AGS细胞凋亡的影响

Dexosome from dendritic cells treated with heat-CO2 pneumoperitoneum induced apotosis of human gastric adenocarcinoma AGS cells

目的 观察热CO2处理后树突状细胞（DC）来源外泌体（DEXs）对胃癌AGS细胞凋亡的影响,探讨腹腔镜胃癌根治术时引入热CO2处理的可行性.方法 建立热CO2气腹体外实验模型,分组处理DC2.4细胞后,提取DEXs并通过透射电镜和Western blot鉴定.将各组DEXs干预AGS细胞后,以细胞计数试剂盒（CCK-8）法检测AGS细胞增殖,膜联蛋白V（Annexin V）/碘化丙锭（PI）双染流式细胞术检测细胞凋亡率,Hoechst 33258染色荧光显微镜检测细胞核形态,比色法检测半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3的活性.结果 成功提取DEXs,热CO2气腹组DEXs可显著抑制胃癌AGS细胞的增殖,l、2、3、4d后抑制率分别为31.13％、43.28％、51.34％和50.32％.细胞出现核固缩、核碎裂和凋亡小体形成等凋亡形态,流式细胞检测表明各组DEXs干预AGS细胞后凋亡率分别为2.3％、5.6％、15.7％和36.8％.与对照组比较,热CO2气腹组Caspase-3相对活性为171.37％.结论 热CO2处理DC后,DEXs能够显著抑制胃癌AGS细胞的增殖并诱导其凋亡,其原因可能与Caspase-3分子活化有关.

Objective To observe the effects of heat-CO2-treated dendritic cells （DC）-derived dexosomes （DEXs） on apoptosis of human gastric adenocarcinoma cell line AGS and investigate the applied value of heat-CO2 pneumoperitoneum during laparoscopic surgery.Methods An in vitro heat-CO2 pneumoperitoneum experimental box was built.After heat-CO2 pneumoperitoneum treatment,the DC-derived DEXs were isolated and identified through TEM and Western blotting.AGS cells were co-cultured with DEXs,apoptosis was examined by Annexin V/propidium iodide （PI） flow cytometry and Hoechst 33258 fluorescent microscopy,and relative Caspase-3 activity was determined by using colorimetric assay.Results DEXs were successfully isolated.Heat-CO2-treated DC-derived DEXs produced evident anti-proliferative effect and the cell proliferation inhibition rate at 1st,2nd,3rd and 4th day was 31.13％,43.28％,51.34％ and 50.32％ respectively.The heat-CO2-treated CD-derived DEXs induced apoptosis of AGS cells.The early apoptosis rate induced by DEXs was 2.3％,5.6％,15.7％ and 36.8％ respectively.The relative viability of AGS cells in heat-CO2-treated DC-derived DEXs group was 171.37％.Conclusion Heat-CO2 treated DC-derived DEXs could significantly inhibit proliferation and induce apoptosis of AGS cells,which may be related to the activation of Caspase-3.