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背景与目的:苯醌安莎霉素类如格尔德霉素(geldanamycin,GA)通过抑制热休克蛋白(heat shock protain90,HSP90)从而导致细胞内信号调节通路网络的多点阻断,如导致涉及细胞增殖、细胞周期调节及幸存调节等众多效应蛋白的降解。前期研究证明HSP90可作为抗肿瘤治疗的一个分子靶位。本研究以子宫颈癌HeLa细胞为研究对象,观察HSP90分子伴侣复合物阻止剂格尔德霉素能否增强加热对HeLa细胞的杀伤效应。方法:采用四唑盐比色实验(MTT)、流式细胞术(flow cytometry,FCM)、吖啶橙荧光染色等方法观察比较应用单纯热疗组、GA组、热疗联合GA组对HeLa细胞活力、凋亡率及细胞形态学的影响。结果:四唑盐比色实验观察到加热与GA化疗间有交互作用,加热联合GA组显著增强单独加热对HeLa细胞杀伤效应(P<0.01);Annexin V-FITC染色流式细胞术结果显示加热联合GA组、单纯加热组、单纯GA治疗组所致细胞凋亡率分别为92.1%、52.2%及26.8%。用吖啶橙荧光染色法观察到加热联合GA组明显见HeLa细胞固缩着色不均而且较深,细胞核浓缩、裂解等凋亡特征。结论:HSP90分子伴侣复合物阻止剂GA能增强热诱导的HeLa细胞的杀伤作用。
BACKGROUND & OBJECTIVE: Benzoquinone ansamycins, such as geldanamycin (GA), inhibit the heat shock protein 90 (HSP90) and lead to multipoint blockade in the intracellular signaling pathway. As a result, Involved in cell proliferation, cell cycle regulation and survival regulation and many other effects of protein degradation. Preliminary studies have shown that HSP90 can be used as a molecular target of anti-tumor therapy. In this study, cervical cancer HeLa cells as the research object, observe HSP90 molecular chaperone antagonist geldanamycin can enhance the heating of HeLa cells killing effect. Methods: MTT, flow cytometry (FCM) and acridine orange staining were used to observe the effects of heat hyperthermia, GA, hyperthermia combined with GA on HeLa cells Vitality, apoptosis rate and cell morphology. Results: The tetrazolium salt colorimetric assay showed that there was interaction between heating and GA chemotherapy. Heating combined with GA group significantly enhanced the killing effect of HeLa cells alone (P <0.01). The results of Annexin V-FITC staining showed that heating The apoptosis rate in combined GA group, simple heating group and simple GA treatment group was 92.1%, 52.2% and 26.8% respectively. Fluorescence staining with acridine orange was observed in the heating combined with GA obviously HeLa cell shrinkage uneven and deep, nuclear condensation, lysis and other apoptosis characteristics. Conclusion: HSP90, a molecular chaperone inhibitor, can enhance the cytotoxicity of heat-inducible HeLa cells.