三氧化二砷抑制DNA损伤修复的放射增敏  

Arsenic trioxide achieves radiosensitization by inhibiting DNA damage repair

作　　者：高兴兴 惠慧 任洪荣 周云[1,2] GAO Xingxing;HUI Hui;REN Hongrong;ZHOU Yun(Department of Affiliated The First Clinical Medical College of Xuzhou Medical University,Xuzhou 221000 China;Xuzhou Central Hospital,Xuzhou 221000 China)

机构地区：[1]徐州医科大学附属第一临床医学院,江苏徐州221000 [2]徐州市中心医院,江苏徐州221000

出　　处：《中国辐射卫生》2024年第4期426-434,共9页Chinese Journal of Radiological Health

基　　金：江苏省妇幼健康项目(F202035),难治型妇科恶性肿瘤SBRT治疗。

摘　　要：目的探讨三氧化二砷(ATO)在宫颈癌中的放射增敏作用,并进一步探索其相关的DNA损伤修复机制。方法体外培养人宫颈癌细胞(Siha、Hela细胞),给予不同浓度的ATO处理,利用CCK-8实验检测细胞增殖情况;将细胞分为4个组:对照(Control)组、放疗(IR)组、ATO组、放疗+给药(IR+ATO)组,平板克隆实验计算放射增敏比;流式细胞术检测细胞周期及凋亡情况;免疫荧光检测γH2AX的表达;蛋白免疫印记法(Western blot)检测Cyclin B1、PTEN、RAD51的表达水平;逆转录聚合酶链式反应(qPCR)检测RAD51的mRNA水平。结果CCK-8结果显示1μM及更高浓度的ATO可以显著抑制Siha及Hela细胞增殖。平板克隆结果显示,ATO对宫颈癌具有放射增敏作用,放射增敏比(SER)分别为1.37、1.30。流式细胞术结果显示放疗+给药组的细胞周期阻滞比例较对照组显著增高(P<0.001);放疗+给药组的凋亡率较对照组明显升高(P<0.01)。蛋白免疫印迹法结果显示放疗+给药组PTEN、RAD51蛋白的表达水平降低(P<0.05)、Cyclin B1蛋白的表达水平升高(P<0.05),差异具有统计学意义。结论ATO通过消耗PTEN来阻滞DNA同源重组修复途径,从而在宫颈癌中达到放射增敏的效果。Objective To explore the radiosensitizing effect of arsenic trioxide(ATO)in cervical cancer,and to further explore the underlying mechanism related to DNA damage repair.Methods Human cervical cancer cells(Siha and Hela cells)were cultured in vitro,treated with different concentrations of ATO,and the cell proliferation was detected by CCK-8 assay.The cells were divided into four groups:control group,radiotherapy(IR)group,ATO group,and radiotherapy+ATO(IR+ATO)group.Radiosensitization ratio was determined by plate cloning assay,cell cycle and apoptosis by flow cytometry,the expression ofγH2AX by immunofluorescence,the expression of Cyclin B1,PTEN,and RAD51 by Western blot,and the expression of RAD51 mRNA by reverse transcription quantitative polymerase chain reaction(RT-qPCR).Results CCK-8 showed that ATO at concentrations of 1μM and higher could significantly inhibit the proliferation of Siha and HeLa cells.Plate cloning showed that ATO had a radiosensitizing effect on cervical cancer,and the radiosensitization ratios were 1.37 and 1.30,respectively.Flow cytometry showed that the proportion of cell cycle arrest was significantly higher in the IR+ATO group than that in the control group(P<0.001).The apoptosis rate was significantly higher in the IR+ATO group than that in the control group(P<0.01).Western blotting showed that the expression levels of PTEN and RAD51 proteins significantly decreased(P<0.05)and the expression level of Cyclin B1 protein significantly increased(P<0.05)in the IR+ATO group.Conclusion ATO achieves radiosensitization in cervical cancer through blocking the DNA homologous recombination repair pathway by consuming PTEN.

关 键 词：三氧化二砷 宫颈癌 DNA损伤修复 放射增敏 

分 类 号：X591[环境科学与工程—环境工程]