槲皮素协同影响DNA修复和ROS水平提高NSCLC细胞的放疗敏感性  

Quercetin Affects Both DNA Repair and ROS Level to Improve Radiosensitivity of NSCLC Cells

作　　者：李怡[1] 陈其章[1] 范临夏[1] 缪国英[2] LI Yi;CHEN Qizhang;FAN Linxia;MIAO Guoying(Department of Respiration in Cadre Ward,Gansu Provincial Hospital,Lanzhou 730000;Department of Radiation Oncology,Gansu Provincial Hospital,Lanzhou 730000)

机构地区：[1]甘肃省人民医院干部呼吸科,兰州730000 [2]甘肃省人民医院放疗科,兰州730000

出　　处：《中药药理与临床》2023年第11期62-67,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：甘肃省自然科学基金(编号:21JR1RA033);甘肃省人民医院科技支撑项目(编号:ZX-62000001-2021-266)。

摘　　要：目的:探究槲皮素对非小细胞肺癌(Non-small cell lung cancer,NSCLC)细胞放疗敏感性的影响及潜在机制。方法:细胞分为空白对照组、单纯辐照组、槲皮素10μmol/L组、槲皮素10μmol/L+辐照组。用MTT法检测槲皮素对细胞的毒性;克隆形成法检测细胞存活率;Western blot法检测Cdc2、p-Cdc2、Cyclin B1、Cdc25c和p-Cdc25c蛋白表达;流式细胞术检测细胞周期和细胞凋亡;活性氧(ROS)和DNA损伤分别用CM-H2DCFDA和免疫荧光法检测。结果:MTT结果表明,10μmol/L的槲皮素对两株细胞的抑制率均在5.0%以内,因此选择该浓度药物做后续研究;10μmol/L的槲皮素预处理显著提高了A549和H1650细胞的辐射敏感性;槲皮素显著上调p-Cdc25c、p-Cdc2和下调Cyclin B1蛋白表达,显著升高G2/M期占比(P<0.01);与单纯辐照组相比,联合处理组DNA双链断裂程度、ROS水平、细胞凋亡率显著升高(P<0.01)。结论:槲皮素通过协同影响DNA修复和ROS水平提高NSCLC细胞的放疗敏感性,有望作为一种增强NSCLC治疗疗效的放射增敏剂。Objective:To explore the effect of quercetin on the radiosensitivity of non-small cell lung cancer(NSCLC)cells and the underlying mechanism.Methods:The cells were classified into control(DMSO),quercetin(10μmol/L),simple irradiation(DMSO+IR),and combined treatment(quercetin 10μmol/L+IR)groups.Methyl thiazolyl tetrazolium(MTT)assay was used to detect the cytotoxicity of quercetin.The colony formation assay was employed to examine the cell survival rate,and western blotting to determine the protein levels of Cdc2,p-Cdc2,Cyclin B1,Cdc25c,and p-Cdc25c.Cell cycle and apoptosis were detected by flow cytometry.The reactive oxygen species(ROS)and DNA damage were examined by CM-H_2DCFDA and immunofluorescence,respectively.Results:The results of MTT assay demonstrated that the inhibition rates of 10μmol/L quercetin on both cell lines were less than 5.0%,and thus 10μmol/L quercetin was selected for the follow-up experiments.The pretreatment with 10μmol/L quercetin significantly increased the radiosensitivity of A549 and H1650 cells.Quercetin induced G2/M phase arrest by up-regulating the expression of p-cdc25c and p-cdc2 and down-regulating cyclin B1,thus increasing the proportion of cells in the G2/M phase(P<0.01).Compared with the simple irradiation group,the combined treatment increased the degree of DNA double-strand break,reactive oxygen species(ROS)level,and apoptosis rate(P<0.01).Conclusion:Quercetin affects both DNA repair and the ROS level to improve the radiosensitivity of NSCLC cells,being expected to serve as a radiosensitizer to enhance the therapeutic efficacy of NSCLC cells.

关 键 词：槲皮素 非小肺癌细胞 放射敏感性 DNA修复 活性氧 

分 类 号：R285[医药卫生—中药学]