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目的探讨热疗中热休克蛋白90(HSP90)对26S蛋白酶体的调控机制。方法建立42℃热疗的Hep G2肝癌细胞系模型,评价不同热疗持续时间(0、3、6、12、24 h)对细胞内活性氧簇(ROS)和细胞增殖的影响;并对热疗导致Hsp90α和26S蛋白酶体的影响进行分析。结果热疗后细胞内ROS的含量增加(F=28.958,P<0.001),且热疗对细胞的增殖抑制程度随着时间的延长显著增强(F=621.704,P<0.001);热疗导致胞内Hsp90α表达量增加(F=27.403,P=0.035),26S蛋白酶体表达量明显降低(F=164.174,P<0.001),活性也下降(F=133.043,P<0.001);干扰HSP90α后,26S蛋白酶体也减少(F=180.231,P<0.001)。结论随着热疗时间延长,细胞内ROS含量增加,热应激和ROS共同导致蛋白持续变性而消耗HSP90,使没有足够的HSP90对26S蛋白酶体组装和稳定导而导致变性蛋白蓄积发生未折叠蛋白反应使细胞死亡。
Objective To investigate the regulation mechanism of 26S proteasome by heat shock protein 90 (HSP90) in hyperthermia. Methods Hep G2 hepatocarcinoma cell line model was established by 42 ℃ hyperthermia. The effects of different hyperthermia duration (0,3,6,12,24 h) on reactive oxygen species (ROS) and cell proliferation were evaluated. The effects of Hsp90α and 26S proteasomes on therapies were analyzed. Results The content of intracellular ROS increased after hyperthermia (F = 28.958, P <0.001), and the degree of inhibition of hyperthermia on cell proliferation increased significantly with time (F = 621.704, P <0.001) (F = 27.403, P = 0.035). The expression of 26S proteasome was significantly decreased (F = 164.174, P <0.001) and the activity of Hsp90α was also decreased (F = 133.043, P <0.001) The proteasome also decreased (F = 180.231, P <0.001). Conclusion With the prolongation of hyperthermia, the content of intracellular ROS increases, heat stress and ROS together lead to the continuous denaturation of proteins and the depletion of HSP90, so that there is not enough HSP90 on 26S proteasome assembly and stabilization of lead and lead to the accumulation of denatured protein unfolded protein The reaction killed the cells.