SDT联用安罗替尼对非小细胞肺癌的协同杀伤作用  

Synergistic cytotoxic effect of sonodynamic therapy combined with anlotinib on non-small cell lung cancer

作　　者：雷月 朱丽全 刘鹏 王浩强 谢波 Yue Lei;Liquan Zhu;Peng Liu;Haoqiang Wang;Bo Xie(Graduate School,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Department of Oncology,General Hospital of the Southern Theater Command,Guangzhou 510010,China)

机构地区：[1]广州中医药大学研究生院,广州市510006 [2]中国人民解放军南部战区总医院肿瘤科

出　　处：《中国肿瘤临床》2024年第20期1027-1033,共7页Chinese Journal of Clinical Oncology

摘　　要：目的:探究声动力疗法(sonodynamic therapy,SDT)联合安罗替尼对非小细胞肺癌(non-small cell lung cancer,NSCLC)的协同杀伤效果及作用机制。方法:本研究选取A549和H1299肺癌细胞作为研究对象,设置空白对照组、单药安罗替尼组、单纯SDT组及SDT联合安罗替尼治疗组。通过CCK-8和细胞划痕实验检测细胞活性与细胞迁移能力;流式细胞术检测活性氧(reactive oxygen species,ROS)水平、凋亡状态和细胞周期;蛋白质印迹法(Western blot)检测凋亡蛋白Caspase-3和周期蛋白Cyclin D1的表达;ROS清除实验等探讨SDT联合安罗替尼协同作用机制。结果:与单纯使用安罗替尼相比,SDT联合安罗替尼可显著抑制肺癌细胞的增殖[细胞存活率:A549:(49.96±4.82)%vs.(86.79±2.64)%,P<0.01;H1299:(31.91±4.87)%vs.(88.04±2.16)%,P<0.001]与迁移[愈合率:A549:(4.23±0.17)%vs.(14.28±0.05)%,P<0.05;H1299:(13.68±2.16)%vs.(42.81±8.11)%,P<0.001]。联合治疗组可明显诱导A549细胞凋亡[凋亡率:(12.58±0.815)%vs.(8.43±0.56)%,P<0.05]。机制研究发现,安罗替尼耐药与ROS水平相关,经活性氧清除剂N-乙酰半胱氨酸(NAC)作用后,细胞内ROS含量减少,安罗替尼IC50增大,敏感性下降。SDT联合安罗替尼后,肿瘤细胞内的ROS水平显著高于单药安罗替尼组(934.14±2.01 vs.166.75±1.45,P<0.001),并激活Caspase-3,下调Cyclin D1的表达。结论:SDT联合安罗替尼具有明显的协同抗肿瘤作用,其机制与ROS通路激活下游凋亡蛋白Caspase-3及下调Cyclin D1,抑制肺癌细胞增殖,诱导细胞凋亡相关。Objective:To investigate the anti-tumor effects and mechanisms of sonodynamic therapy(SDT)combined with anlotinib on nonsmall cell lung cancer(NSCLC).Methods:A549 and H1299 cells were used as the research models and the following groups were established:control,anlotinib,SDT,and SDT-anlotinib.Cell viability and migration ability were assessed using the CCK-8 and cell scratch assays.Additionally,flow cytometry was employed to determine reactive oxygen species(ROS)levels,apoptosis,and cell cycle;Western blot was performed to detect Caspase-3 and Cyclin D1 expression;and ROS elimination experiments were conducted to explore the mechanisms of combined SDT and anlotinib treatment.Results:In comparison to the anlotinib group,the SDT-anlotinib group demonstrated a notable inhibition in the proliferation of lung cancer cells[cell viability:A549:(49.96±4.82)%vs.(86.79±2.64)%,P<0.01;H1299:(31.91±4.87)%vs.(88.04±2.16)%,P<0.001]and migration[healing rate:A549:(4.23±0.17)%vs.(14.28±0.05)%,P<0.05;H1299:(13.68±2.16)%vs.(42.81±8.11)%,P<0.001].Furthermore,the combination therapy group exhibited a notable induction of apoptosis[apoptosis rate:(12.58±0.815)%vs.(8.43±0.56)%,P<0.05].Mechanistic studies have demonstrated that anlotinib resistance is associated with ROS levels.Treatment with the ROS scavenger N-acetylcysteine(NAC)has been shown to decrease intracellular ROS content,increase the IC50 of anlotinib,and reduce sensitivity.Intracellular ROS levels in tumor cells were significantly higher in SDT-anlotinib group compared to that in the anlotinib group[(934.14±2.01)vs.(166.75±1.45),P<0.001].Additionally,Caspase-3 activation was observed,accompanied by a reduction in Cyclin D1 expression.Conclusions:The combination of SDT and anlotinib exerted a pronounced anti-tumor effect.Activation of the ROS pathway led to the activation of Caspase-3 and the downregulation of Cyclin D1,resulting in the inhibition of lung cancer cell proliferation and the induction of apoptosis.

关 键 词：非小细胞肺癌 声动力疗法 安罗替尼 ROS 

分 类 号：R734.2[医药卫生—肿瘤]