【摘要】 目的:探讨Tat-SmacN7融合肽对人非小细胞肺癌H460细胞辐射增敏的作用及其机制。方法:WST-1法检测Tat-SamcN7对H460细胞的毒性作用,细胞克隆实验观察Tat-SamcN7对H460细胞的辐射增敏作用,流式细胞术检测H460细胞的凋亡率,ELISA法检测H460细胞的Caspase-3、Caspase-8和Caspase-9的活性。结果:0.01~100μmol/L的Tat-SmacN7融合肽与H460细胞共培养,细胞存活率均>80%;20μmol/L的Tat-SmacN7融合肽与H460细胞共培养,对细胞的辐射增敏比为1.63;Tat-SmacN7(20μmol/L)对细胞凋亡〔(10.87±0.45)%〕有主效应,F=54.389,P<0.001;4Gy照射对细胞凋亡〔(19.83±0.75)%〕有主效应,F=641.516,P<0.001;二者有交互效应,F=65.756,P<0.001。Tat-SmacN7对细胞Caspase-3活性(1.418±0.064)有主效应,F=67.029,P<0.001;照射对细胞Caspase-3活性(1.188±0.102)有主效应,F=29.509,P=0.001;两者有交互作用,F=11.328,P=0.01。Tat-SmacN7对细胞Caspase-8活性(1.867±0.208)有主效应,F=149.705,P<0.001;照射对细胞Caspase-8活性(1.227±0.123)有主效应,F=30.418,P=0.001;两者有交互作用,F=10.663,P=0.011。Tat-SmacN7对细胞Caspase-9活性(1.672±0.075)有主效应,F=127.027,P<0.001;照射对细胞Caspase-9活性(1.279±0.141)有主效应,F=44.281,P<0.001;两者有交互作用,F=9.782,P=0.014。结论:Tat-SmacN7单独作用对非小细胞肺癌H460细胞没有明显毒性作用,但能增强H460细胞对辐射的敏感性,具有辐射增敏作用,与其能增强细胞内Caspase-3、Caspase-8和Caspase-9的活性相关。更多还原

【Abstract】 OBJECTIVE:To discover radiosensitization of Tat-SmacN7fusion peptide to human non-small cell lung cancer cell line H460and its mechanism.METHODS:The WST-1assay was used to detect toxicity of Tat-SamcN7to H460cell line;Clonogenic assay was done to examine the radiosensitization of Tat-SamcN7to H460;Flow cytometry were used to examine the apoptosis of H460cell line.Activation of Caspase-3,Caspase-8and Caspase-9was detected by Caspase activity assays based on ELISA.RESULTS:Tat-SmacN7fusion peptide with concentration range of 10 nmol/L to 100μmol/L was co-cultured with H460 cells,cell survival rates were 80% or more;Tat-SmacN7fusion peptide(20μmol/L)and H460cells were co-cultured,the cell radiosensitizing ratio was 1.63;Tat-SmacN7(20μmol/L)had main effect to cell apoptotic rate[(10.87±0.45)%,F=54.389,P<0.001];Radiation had main effect to cell apoptotic rate[(19.83±0.75)%,F=641.516,P<0.001];They had interaction(F=65.756,P<0.001).Tat-SmacN7had main effect on the activity of Caspase-3(1.418±0.064,F=67.029,P<0.001);Radiation had main effect on activity of Caspase-3(1.188±0.102,F=29.509,P=0.001);They had interaction(F=11.328,P=0.010).Tat-SmacN7had main effect on activity of Caspase-8(1.867±0.208,F=149.705,P<0.001);Radiation had main effect on activity of Caspase-8(1.227±0.123,F=30.418,P=0.001);They had interaction(F=10.663,P=0.011).Tat-SmacN7had main effect on activity of Caspase-9(1.672±0.075,F=127.027,P<0.001);Radiation had main effect on activity of Caspase-9(1.279±0.141,F=44.281,P<0.001);They had interaction(F= 9.782,P=0.014).CONCLUSION:Tat-SmacN7does not have toxicity to H460cell line used as a single agent,but can increase the sensitivity of H460cell to radiation,and the radiosensitization have relation with the increase of activity of Caspase-3,Caspase-8and Caspase-9.更多还原