安罗替尼通过抑制PI3K/Akt/ERK通路增强鼻咽癌CNE-2细胞放疗敏感性的机制研究OA
Mechanism study of anlotinib enhancing radiosensitivity of nasopharyngeal carcinoma CNE-2 cells by inhibiting the PI3K/Akt/ERK pathway
目的 探讨安罗替尼对鼻咽癌CNE-2细胞放疗敏感性的影响及其可能的作用机制.方法 将CNE-2细胞分为对照组、安罗替尼组、放疗组及联合组(安罗替尼联合放疗).采用 MTT法检测不同浓度安罗替尼对CNE-2细胞增殖的影响;Western blot检测放疗(X射线照射)及安罗替尼处理对磷酸肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/细胞外信号调节激酶(ERK)通路磷酸化水平、凋亡相关蛋白[cleaved半胱天冬酶-3(caspase 3)、髓细胞白血病-1(Mcl-1)]及DNA损伤标志物磷酸化组蛋白H2AX(γ-H2AX)表达的影响;流式细胞术检测细胞周期分布;克隆形成实验评估安罗替尼对放疗敏感性的影响.结果 安罗替尼呈浓度和时间依赖性抑制CNE-2细胞增殖(P<0.05).4 Gy放疗可明显上调磷酸化(p)-PI3K、p-Akt及p-ERK蛋白表达水平(P<0.05),而安罗替尼可有效抑制上述磷酸化蛋白表达水平(P<0.05).与对照组比较,联合组G2/M期细胞比例增加25.7%,S期比例下降22.4%(P<0.05);cleaved caspase 3蛋白表达水平明显升高,Mcl-1蛋白表达水平明显下降(P<0.05).联合组γ-H2AX蛋白表达水平明显高于放疗组(P<0.05).克隆形成实验表明,20.0 μmol/L联合组的增敏效应最明显,放疗增敏比最高(P<0.05).结论 安罗替尼可以增加鼻咽癌细胞放疗敏感性,其机制可能与抑制PI3K/Akt/ERK通路、诱导G2/M期周期阻滞、促进细胞凋亡及阻碍DNA双链断裂修复有关.
Objective To investigate the effect of anlotinib on radiosensitivity of nasopharyngeal carci-noma CNE-2 cells and its potential mechanism.Methods CNE-2 cells were divided into control group,anlo-tinib group,radiotherapy group and combined group(anlotinib combined with radiotherapy).The MTT assay was used to detect the proliferative viability of CNE-2 cells treated with different concentrations of anlotinib.Western-blot was performed to determine the phosphorylation levels of phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/extracellular signal-regulated kinase(ERK)pathway,the expression of apoptosis-re-lated proteins[cleaved caspase 3,myeloid cell leukemia-1(Mcl-1)]and DNA damage marker phosphorylated histone H2AX(γ-H2AX)after X-ray irradiation and anlotinib intervention.Flow cytometry was adopted to analyze cell-cycle distribution,and colony-formation assay was carried out to evaluate the radiosensitizing effect of anlotinib.Results Anlotinib inhibited the proliferation of CNE-2 cells in a concentration-and time-dependent manner(P<0.05).4 Gy irradiation significantly up-regulated the protein levels of phosphorylated(p)-PI3K,p-Akt and p-ERK(P<0.05),whereas anlotinib effectively suppressed the expression of these phosphorylated proteins(P<0.05).Compared with the control group,the proportion of cells in G2/M phase in the combined group increased by 25.7%while the proportion of S-phase cells decreased by 22.4%(P<0.05);the expression of cleaved caspase 3 was markedly elevated and Mcl-1 was obviously down-regulated(P<0.05).The γ-H2AX level in the combined group was significantly higher than that in the radiotherapy group(P<0.05).Colony-formation experiments demonstrated that the 20.0 μmol/L combination group ex-hibited the most prominent sensitization effect with the highest radiosensitization ratio(P<0.05).Conclusion Anlotinib can enhance the radiosensitivity of nasopharyngeal carcinoma cells.The underlying mechanism may involve inhibiting the PI3K/Akt/ERK signaling pathway,inducing G2/M-phase cell-cycle arrest,facilita-ting cell apoptosis and impairing the repair of DNA double-strand breaks.
赵明;秦秋子;杨艳萍;高岭
开封市中心医院肿瘤放疗科,河南 开封 475000开封市中心医院影像科,河南 开封 475000开封市中心医院肿瘤放疗科,河南 开封 475000开封市中心医院肿瘤放疗科,河南 开封 475000
医药卫生
鼻咽癌安罗替尼放疗敏感性磷酸肌醇3-激酶/蛋白激酶B通路DNA损伤修复
nasopharyngeal carcinomaanlotinibradiosensitivityphosphoinositide 3 kinase/protein ki-nase B pathwayDNA damage repair
《重庆医学》 2026 (8)
1715-1721,7
河南省医学科技攻关计划项目(LHGJ20240384).
评论