DC-exosomes联合顺铂对人胶质瘤U251细胞增殖和凋亡的影响

摘要

Objective To observe the effect of cisplatin combinated with the therapeutic DC-exosomes vaccine on U251 glioma cells proliferation and apoptosis. Methods The U251 cells were divided into four groups randomly. Cells in group A were cultured with conventional nutrient solution, activated lymphocyte by DC-exosomes (2 ×106/mL)0. 5 mL to B, cisplatin ( 1 mg/mL) 0. 01 mL to C, activated lymphocyte by DC-exosomes ( 2 × 10 /mL)0. 5 mL and cisplatin ( 1 mg/ mL) 0.01 mL to D, and train those cells 2 d. The proliferation rate of U251 were detected by MTT and the apoptosis by the flow cytometry. Result After cultured with MTT for 12,24,36,48,60 h,the OD of goroup A was respectively 1. 38 ± 0.01,1.36 ±0.02,1.34 ±0. 03,1.32 ±0.05,1.30 ±0.03; the OD of goroup B was respectively 1. 30 ±0. 02,1. 10 ± 0.03,0.90 ±0.04,0.75 ±0.03,0.62 ±0.03; the OD of goroup C was respectively 1. 00 ±0. 04,0. 80 ±0. 02,0. 55 ± 0.04,0.30 ±0.03,0. 15 ±0.04,while the OD of goroup D was respectively 0.75 ±0.04,0.50 ±0.04,0.38 ±0.02,0.21 ±0.02,0.05 ±0.03. The OD of group A were higher than that in group B,C,D (all P<0.05) , the OD of group B and C were higher than thatin group D( all P < 0. 05 ) . After the training of 24 h, the cell apoptosis rate of group A, B, C, D were 0.5% ±0.3%,18.6% ±1.7% ,36.9% ±4.3% ,57.5% ±2.1% respectively. The cell apoptosis rate of group A were higher than that in group B,C,D ( all P <0. 05), the cell apoptosis rate of of group B and C were higher than thatin group D ( all P < 0. 05 ) . Conclusion DC-exosomes vaccine combined with cisplatin can restrain the proliferation of U251 glioma and also lead to the apoptosis of U251 cells.%目的 观察利用人树突细胞(DC)制备的治疗性疫苗DC-exosomes联合顺铂对人脑胶质瘤U251细胞增殖与凋亡的影响.方法 将传代的U251细胞随机分为四组,A组加常规培养液,B、C、D组分别加DC-exosomes激活的淋巴细胞(2×106/mL)0.5 mL、顺铂(1 mg/mL)0.01 mL、DC-exosomes激活淋巴细胞(2×106/mL)0.5 mL+顺铂(1 mg/mL)0.01 mL培养2d；用MTT法检测U251细胞光密度值(OD值)观察细胞增殖情况,用流式细胞仪检测U251细胞凋亡率.结果 加入MTT培养12、24、36、48、60 h,A组细胞OD值分别为1.38 ±0.01、1.36±0.02、1.34±0.03、1.32 ±0.05、1.30±0.03,B组分别为1.30±0.02、1.10 ±0.03、0.90±0.04、0.75±0.03、0.62 ±0.03,C组分别为1.00±0.04、0.80±0.02、0.55 ±0.04、0.30±0.03、0.15±0.04,D组分别为0.75±0.04、0.50±0.04、0.38±0.02、0.21±0.02、0.05±0.03；B、C、D组与A组比较,P均＜0.05；D组与B、C组比较,P均＜0.05.培养24 h时,A、B、C、D组细胞凋亡率分别为0.5％±0.3％、18.6％±1.7％、36.9％±4.3％、57.5％±2.1％.B、C、D组与A组比较,P均＜0.05；D组与B、C组比较,P均＜0.05.结论 DC-exosomes疫苗联合顺铂可抑制人脑胶质瘤U251细胞增殖,并诱导其凋亡.