17AAG-cypate聚合物胶束对肺癌A549细胞放射敏感性影响

目的 研究17AAG-cypate胶束对人非小细胞肺癌A549细胞的放射增敏作用,并简析其可能机制。方法 单击多靶模型拟合实验方程分析17AAG-M和17AAG-cypate-M的放射增敏作用。MTT实验检测17AAG-cypate-M在激光和X射线照射下对A549细胞存活率的影响。β-半乳糖苷酶染色实验观察药物对细胞衰老产生的影响。γ-H₂AX免疫荧光染色实验分析不同处理条件对DNA损伤修复的影响。蛋白印迹法实验检测p-Erk1/2和p-Akt蛋白的表达情况。配对t检验差异。结果 与单纯X射线照射组相比D₀下降,SER>1,说明17AAG-cypate-M具有放射增敏作用。与17AAG-M组相比,17AAG-cypate-M组对A549细胞活力抑制程度升高(P=0.0012),且引发了更明显的DNA损伤修复延迟(P=0.0017)。同时,17AAG-cypate-M对p-Erk1/2和p-Akt表达水平的抑制程度高于17AAG-M。结论 与17AAG-M相比,17AAG-cypate-M对A549细胞的放射增敏作用更加明显,其放射增敏机制可能为引发细胞的衰老,延迟DNA损伤修复,抑制p-Erk1/2和p-Akt表达等。

Effect of 17AAG-cypate polymer micelle on radio-sensitivityof A549 cells

Objective To investigate the radiosensitizing effect of 17AAG-cypate micelles on human non-small cell lung cancer A549 cells and its possible mechanism.Methods (1) A single-hit multi-target model formula was used to analyze the radiosensitizing effects of 17AAG-M and 17AAG-cypate-M.(2) The effects of 17AAG-cypate-M on the viability of A549 cells under laser and X-ray irradiation were analyzed by MTT assay.(3) The effect of the drugs on the cell senescence was observed by β-galactosidase staining assay.(4) The effects of different treatment conditions on DNA damage repair were analyzed by γ-H2AX immunofluorescence staining assay.(5) The expression of p-Erk1/2 and p-Akt was measured by Western blot.The paired t test was used for analyzing the differences between groups.Results Compared with the X-ray irradiation group,the X-ray+17AAG-cypate-M group had a lower mean lethal dose and a sensitization enhancement ratio greater than 1,indicating that 17AAG-cypate-M had a radiosensitizing effect.Compared with the 17AAG-M group,the 17AAG-cypate-M group showed significantly lower cell viability (P<0.01),a significantly higher percentage of aging cells (P<0.01),and significantly further delayed DNA damage repair (P<0.01).And the 17AAG-cypate-M group had lower expression of p-Erk1/2 and p-Akt than the 17AAG-M group.Conclusions Compared with 17AAG-M,17AAG-cypate-M has a higher radiosensitizing effect on A549 cells.The mechanism might be inducing the cell senescence,delaying DNA damage repair,and inhibiting the expression of p-Erk1/2 and p-Akt.