摘要
目的研究安罗替尼对肺腺癌细胞放疗增敏作用及机制。方法采用安罗替尼处理人肺腺癌细胞株A549,使用CCK8法测定安罗替尼对细胞增殖的影响;采用克隆形成实验测定安罗替尼联合放疗对细胞的生长抑制作用;使用流式细胞术测定细胞周期和细胞凋亡的变化;采用免疫荧光测定安罗替尼联合放疗对细胞内DNA损伤的影响;采用Western blot检测细胞内DNA损伤标志因子DNA-PKcs的表达变化。结果安罗替尼对人肺腺癌细胞A549增殖具有抑制作用,第2、3、4、5天,(P<0.05)。体外培养克隆形成实验显示,安罗替尼联合放疗组的准予剂量(Dq)、平均致死剂量(D0)及4 Gy照射时的存活分数(SF)均明显低于单纯放疗组(P<0.05)。安罗替尼能够使细胞阻滞于G1/G0期,与放疗联合作用后,进一步降低了G2和S期细胞比例。安罗替尼能够增加放疗诱导的细胞凋亡比例(31.94±2.25)%和(44.44±2.30)%,(P<0.001),增加γH2AX阳性细胞数量(25.67±2.52)%和(54.67±3.79)%,(P<0.001)。安罗替尼能够降低放疗诱导的DNA断裂损伤标志因子DNA-PKcs的表达强度(0.90±0.06)和(0.40±0.06),(P<0.001)。结论安罗替尼具有放疗增敏作用,其机制可能与减少G2/S期细胞、增加细胞凋亡和维持DNA持续损伤有关。
Objective To investigate the effect and mechanism of anlotinib on radiosensitization of human lung adenocarcinoma cell line A549. Methods Human lung adenocarcinom cell line A549 was treated with anlotinib and/or radiotherapy,then divided in to four groups,control group(Ctrl),Anlotinib treatment group(A),irradiation group( RT) and anlotinib combined with irradiation group( A + RT). CCK8 method was used to determine cell proliferation;the clone formation experiment was used to determine the inhibitory effect on cell growth;flow cytometry was used to determine cell cycle and apoptosis;immunofluorescence of γ-H2AX was used to determine DNA damage;expression of DNA-PKcs were detected by Western blot. Results Anlotinib inhibited proliferation and clonogenic survival following irradiation. The dose(Dq),the average lethal dose(D0) and the survival score( SF2) in the anlotinib combined radiotherapy group was significantly lower than those in the radiotherapy group. Anlotinib decreased G2/M phase arrest and promoted the cells apoptosis induced by in irradiation. The confocal microscopy results showed the average number of γ-H2 AX foci in the A+RT group was more than that in RT group. The protein levels of DNA-PKcs were higher in A + RT group than that in RT group.Conclusion Anlotinib enhances the radiosensitivity of A549 cells,which may be attributed to the delay DNA damage repair. It provides a rationale strategy by Anlotinib combined with irradiation for NSCLC.
作者
段宏民
陈楠
杨永燕
李云霞
冯金象
李航
黄秀文
吕东津
张明
赵玉涛
DUAN Hong-min;CHEN Nan;YANG Yong-yan;LI Yun-xia;FENG Jin-xiang;LI Hang;HUANG Xiu-wen;LV Dong-jin;ZHANG Ming;ZHAO Yu-tao(Dept.of Oncology,The 8th Affiliated Hospital of Dali University/The People's Hospital of Lincang,Lincang Yunnan 677000;Dept.of Thoracic Surgery,The 3rd Affiliated Hospital of Kunming Medical University/Yunnan Cancer Hospital,Kunming Yunnan 650118;Dept.of Geriatric Medicine,The 3th Affiliated Hospital ofDali University/The People fs Hospital ofLincang,Lincang Yunnan 677000;Dept.of Medical Oncology,The 3rd Affiliated Hospital of Kunming Medical University/Yunnan Cancer Hospital,Kunming Yunnan 650118;Dept.of Radiotherapy Center,The 3rd Affiliated Hospital of Kunming Medical University/Yunnan Cancer Hospital,Kunming Yunnan 650118,China)
出处
《昆明医科大学学报》
CAS
2021年第8期40-46,共7页
Journal of Kunming Medical University
基金
国家自然科学基金资助项目(81860537)
云南省教育厅科学研究基金资助项目(2020J0595,2019J1275)
白求恩医学科学研究基金资助项目(B19400DT)。
关键词
安罗替尼
肺腺癌
放疗增敏
DNA损伤修复
Anlotinib
Non-small cell lung cancer
Radiosensitization
DNA damage repair
