热CO_2处理树突状细胞源性外体在胃癌细胞增殖调控中的作用研究

Study on the role of dexosome from dendritic cells treated with hyperthermic CO_2on proliferation regulation of human gastric cancer cells

目的探讨热CO2处理后树突状细胞(DC)源性外泌体(DEX)在胃癌细胞增殖调控中的作用机制。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DEX并通过透射电镜鉴定其形态。体外培养胃癌AGS细胞,用各组DEX干预后以细胞计数试剂盒(CCK-8)法检测AGS细胞的增殖情况,蛋白免疫印迹法(Western blot)检测AGS细胞中蛋白激酶B(AKT)磷酸化水平和B细胞淋巴瘤-2(bcl-2)的表达,比色法检测p-AKT和bcl-2的活性。结果成功提取DEX,热CO2气腹组AGS细胞的增殖率为54.73±2.04%,与单纯热处理组及对照组比较差异有显著性(P<0.05)。Western blot结果显示,热CO2气腹组AGS细胞中p-AKT和bcl-2蛋白表达均显著降低。与对照组比较,热CO2气腹组p-AKT和bcl-2相对活性为41.28±2.31%和36.49±1.92%。结论热CO2处理DC后,DEX能显著抑制胃癌AGS细胞的增殖,其作用机制可能与抑制AKT通路活化和bcl-2表达及活性的降低有关。

Objective To explore the role of dendritic cells （DC）-derived dexosome （DEX） in proliferation regulation of human gastric adenocarcinoma cells after treated with hyperthermic CO2. Methods Based on an in vitro hyperthermic CO2 pneumoperitoneum experimental model,the DC- derived DEX was isolated and identified through TEM after treatment by hyperthermic CO2. AGS cells were co-cultured with DEX in vitro. And the cell proliferation was detected by CCK-8 assay,the expression of p-AKT and bcl-2 in AGS cells was examined by Western blot, and relative activity of p-AKT and bcl-2 was determined by colorimetric assay. Results DEX was successfully isolated. The proliferation rate of AGS cells was 54.73%~2.04% in hyperthermic CO2 treated group, lower than that of heat treated group and control group （P〈0.05）. Significantly lower expression of p-AKT and bcl-2 in AGS cells treated with hyperthermic CO2-DEX was observed by Western blot. And the relative activity of p-AKT and bcl-2 in AGS cells treated with hyperthermic CO2-DEX were 41.28%±2.31% and 36.49%±1.92%. Conclusions DC-derived DEX treated by hyperthermic CO2 could significantly inhibit proliferation of AGS cells, which may be related to the inhibition of AKT pathway activation and the decrease of expression of bcl-2.