DC-exosomes联合顺铂对人胶质瘤U251细胞增殖和凋亡的影响

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2 d;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。

Influence of DC-exosomes vaccine combined with cisplatin on proliferation and apoptosis in human glioblastoma U251 cells

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(2×106/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.