抑制磷脂酰肌醇-3-羟基激酶/蛋白激酶B信号传导通路对宫颈癌放疗增敏的作用

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目的:探讨抑制磷脂酰肌醇-3-羟基激酶/蛋白激酶B(PI3K/AKT)信号传导通路对宫颈癌的放疗增敏作用。方法:选用LY294002为PI3K阻滞剂。培养PI3K功能活性和AKT磷酸化水平稳定的人宫颈癌HeLa细胞株,皮下注射于BALB/C裸鼠体内,建立裸鼠宫颈癌移植瘤模型。随机将荷瘤裸鼠分为空白对照组、单纯放疗组、单纯药物组(LY294002)和联合治疗组(放疗+LY294002)。应用Western blotting检测各组AKT磷酸化(phospho-Ser473AKT)水平。通过克隆形成实验检测LY294002在体内的放射增敏作用。长期观察肿瘤体积的变化并绘制肿瘤生长曲线。结果:Western blotting显示,联合治疗组的AKT去磷酸化现象最显著。克隆形成实验显示,LY294002与放射治疗对肿瘤细胞生长的抑制在各剂量水平均表现出协同作用(P<0.05)。肿瘤生长曲线表明联合治疗组的肿瘤体积增长最缓慢。结论:PI3K阻滞剂显著增强宫颈癌对放射治疗的敏感性。 Objective: To investigate the inhibition of radiosensitization of cervical cancer by PI3K / AKT signal transduction pathway. Methods: LY294002 was selected as PI3K blocker. The human cervical cancer HeLa cell line with stable PI3K activity and AKT phosphorylation level was cultured and subcutaneously injected into BALB / C nude mice to establish a nude mouse cervical cancer xenograft model. Tumor bearing nude mice were randomly divided into blank control group, radiotherapy alone group, simple drug group (LY294002) and combination therapy group (radiotherapy + LY294002). The level of phospho-Ser473AKT in each group was detected by Western blotting. The radiosensitization of LY294002 in vivo was detected by clonogenic assay. Long-term observation of tumor volume changes and draw the tumor growth curve. Results: Western blotting showed that AKT dephosphorylation in combination therapy group was the most significant. Clone formation experiments showed that LY294002 and radiotherapy inhibited the growth of tumor cells at all dose levels showed synergistic effects (P <0.05). Tumor growth curves showed that the combined treatment group had the slowest increase in tumor volume. Conclusion: PI3K blockers significantly enhance the sensitivity of cervical cancer to radiation therapy.
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