CAJ | 학술논문

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.
목적 관찰열CO2처리후수돌상세포(DC)래원외비체(DEXs)대위암AGS세포조망적영향,탐토복강경위암근치술시인입열CO2처리적가행성.방법 건립열CO2기복체외실험모형,분조처리DC2.4세포후,제취DEXs병통과투사전경화Western blot감정.장각조DEXs간예AGS세포후,이세포계수시제합(CCK-8)법검측AGS세포증식,막련단백V(Annexin V)/전화병정(PI)쌍염류식세포술검측세포조망솔,Hoechst 33258염색형광현미경검측세포핵형태,비색법검측반광안선천동안산특이성단백매(Caspase)-3적활성.결과 성공제취DEXs,열CO2기복조DEXs가현저억제위암AGS세포적증식,l、2、3、4d후억제솔분별위31.13％、43.28％、51.34％화50.32％.세포출현핵고축、핵쇄렬화조망소체형성등조망형태,류식세포검측표명각조DEXs간예AGS세포후조망솔분별위2.3％、5.6％、15.7％화36.8％.여대조조비교,열CO2기복조Caspase-3상대활성위171.37％.결론 열CO2처리DC후,DEXs능구현저억제위암AGS세포적증식병유도기조망,기원인가능여Caspase-3분자활화유관.
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.