抑制IGF-1R对喉癌Hep-2细胞放射增敏作用的机制探讨  

Mechanism of inhibiting the radiosensitization of IGF-1R on laryngeal carcinoma Hep-2 cells

作　　者：赵明 秦秋子 侯欣[3] 弋振营 张贝贝 高岭 ZHAO Ming;QIN Qiuzi;HOU Xin;YI Zhenying;ZHANG Beibei;GAO Ling(Department of Radiation Oncology,Henan Kaifeng 475000,China;Department of Imaging,Kaifeng Central Hospital,Henan Kaifeng 475000,China;The First Affiliated Hospital of Henan University,Henan Kaifeng 475000,China)

机构地区：[1]开封市中心医院放疗科,河南开封475000 [2]开封市中心医院影像科,河南开封475000 [3]河南大学第一附属医院,河南开封475000

出　　处：《现代肿瘤医学》2025年第1期20-25,共6页Journal of Modern Oncology

基　　金：河南省医学科技攻关联合共建项目(编号:LHGJ20210559)。

摘　　要：目的:探讨小分子酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI)Linsitinib对喉癌Hep-2细胞放疗增敏的作用机制。方法:MTT检测Linsitinib对Hep-2细胞增殖的影响;Western blot检测X线照射(irradiation,IR)、Linsitinib、X线照射联合Linsitinib对IGF-1R/AKT和IGF-1R/ERK信号通路的影响;流式细胞仪检测Linsitinib对细胞周期的影响;Western blot检测凋亡相关蛋白表达;细胞克隆形成实验分析Linsitinib对X线照射敏感性的作用。Western blot检测细胞DNA损伤相关蛋白γ-H2AX的表达水平。结果:Linsitinib明显抑制Hep-2细胞增殖;X线照射可以激活IGF-1R/AKT和IGF-1R/ERK信号通路,Linsitinib可以抑制IGF-1R/AKT和PI3K/ERK信号通路激活,而Linsitinib联合X线照射进一步抑制IGF-1R/AKT和PI3K/ERK信号通路激活;细胞周期分析显示,Linsitinib可以增加细胞G_(2)/M期比例;Linsitinib联合X线照射可以提高促凋亡蛋白Bax的表达水平,而降低抑凋亡蛋白Bcl-xl的表达水平。细胞克隆形成实验结果提示Linsitinib可提高X线照射对细胞的放射敏感性;Western blot实验提示Linsitinib增加X线照射细胞的γ-H2AX蛋白表达水平。结论:Linsitinib增加喉癌细胞放疗敏感性,其机制可能与抑制PI3K/AKT和Ras/MAPK细胞增殖信号通路激活,改变细胞周期,促进细胞凋亡,诱导基因组不稳定性有关。Objective:To explore the mechanism of tyrosine kinase inhibitor(TKI)Linsitinib enhance the radiosensitivity in larngeal carcinoma cell line.Methods:We examined the effect of Linsitinib on cell proliferation by MTT test.Then,the activation of AKT and ERK1/2 which were downstream of IGF-1R,were detected after X-ray irradiation(IR)and Linsitinib addition by Western blot.The cell cycle was examined by flow cytometry.The expression of Bax,Bcl-xl protein was assayed by Western blot.The survival fraction of Hep-2 was measured by clonogenic assay.Finally,Western blot analysis was performed to detect the expression level of DNA damage-related proteinγ-H2AX.Results:Linsitinib,significantly suppressed Hep-2 cell proliferation.X-ray radiation can activate IGF-1R/AKT and IGF-1R/ERK signaling pathways,but both IGF-1R/AKT and IGF-1R/ERK signaling pathways were suppressed by Linsitinib,but Linsitinib combined with IR further inhibited the activation of IGF-1R/AKT and IGF-1R/ERK signaling pathways.Linsitinib induced Hep-2 cell cycle arrest in G_(2)/M phase.Linsitinib combined with IR can increase the expression level of pro-apoptotic protein Bax and decrease the expression level of anti-apoptotic protein Bcl-xl.The results of cell cloning suggested that Linsitinib increased the radiosensitivity of IR cells.Western blot assay showed that Linsitinib increased the expression ofγ-H2AX protein in X-ray irradiated cells.Conclusion:Linsitinib may increase the radiotherapy sensitivity of laryngeal cancer cells,and its mechanism may be related to inhibiting the activation of PI3K/AKT and Ras/MAPK cell proliferation signaling pathway,changing cell cycle,promoting cell apoptosis,and inducing genomic instability.

关 键 词：喉癌 放射治疗 胰岛素样生长因子受体 酪氨酸激酶抑制剂 放射敏感性 

分 类 号：R739.6[医药卫生—肿瘤]