褪黑素对尼莫司汀抑制胶质瘤U118细胞增殖的增敏效应研究  

Sensitizing effect of melatonin on nimustine in inhibiting the proliferation of glioma U118 cells

作　　者：龙艳春 朱佳斌 孙欢[1] 郑旭 尚发军[1] LONG Yan-chun;ZHU Jia-bin;SUN Huan;ZHENG Xu;SHANG Fa-jun(Department of Neurosurgery,Affiliated Renhe Hospital of China Three Gorges University,Yichang 443000,Hubei,CHINA)

机构地区：[1]三峡大学附属仁和医院神经外科,湖北宜昌443000

出　　处：《海南医学》2022年第7期817-820,共4页Hainan Medical Journal

基　　金：湖北省宜昌市医疗卫生研究项目(编号:A20-2-046)。

摘　　要：目的观察褪黑素对尼莫司汀抑制胶质瘤U118细胞增殖的增敏效应并探讨其机制。方法褪黑素和尼莫司汀单独使用或者联合使用处理胶质瘤U118细胞,采用CCK-8实验检测细胞的活性。将细胞分为对照组、褪黑素组(0.75 mmol/L)、尼莫司汀组(100μmol/L)、联合治疗组(0.75 mmol/L褪黑素联合100μmol/L尼莫司汀),采用协同实验、平板克隆实验、RT-qPCR实验、western blot实验检测褪黑素对尼莫司汀抑制胶质瘤U118细胞的增敏效应及其机制。结果与对照组比较,褪黑素、尼莫司汀单独或者协同给药对胶质瘤U118细胞均具有生长抑制作用,差异均有统计学意义(P<0.05);协同实验中,褪黑素联合尼莫司汀时,U118细胞的两药相互作用指数(coefficient of drug interaction,CDI)为0.43,提示两药有协同抑制作用;平板克隆实验中,褪黑素联合尼莫司汀后U118细胞克隆形成率明显减少,差异有统计学意义(P<0.05);RT-qPCR实验中,褪黑素联合尼莫司汀后,U118细胞中COX-2m RNA的表达明显下调,差异有统计学意义(P<0.05);Western blot实验中,褪黑素联合尼莫司汀后,U118细胞中COX-2蛋白、p-IκBα/IκBα表达明显下调,差异均有统计学意义(P<0.05)。结论褪黑素可增强尼莫司汀对胶质瘤U118细胞增殖的抑制作用。Objective To observe the sensitizing effect of melatonin on nimustine in inhibiting the proliferation of glioma U118 cells,and to explore its mechanism.Methods Glioma U118 cells were treated with melatonin and nimustine alone or in combination,and the cell viability was detected by Cell Counting Kit-8(CCK-8).The cells were divided into control group,melatonin group(0.75 mmol/L),nimustine group(100μmol/L),combined treatment group(0.75 mmol/L melatonin combined with 100μmol/L nimustine).The synergistic experiment,plate cloning experiment,RT-qPCR experiment,western blot experiment were used to detect the sensitizing effect of melatonin on nimustine’s inhibition of glioma U118 cells and its mechanism.Results Compared with the control group,melatonin and nimustine alone or in combination showed inhibitory effect on glioma U118 cells,and the difference was statistically significant(P<0.05).In the synergistic experiment,in combined treatment group,the coefficient of drug interaction(CDI)of U118 cells was 0.43,suggesting that the two drugs have a synergistic inhibitory effect.In plate cloning experiment,after melatonin combined with nimustine,the U118 cell clone formation rate was significantly reduced,and the difference was statistically significant(P<0.05).In the RT-qPCR experiment,after melatonin combined with nimustine,the expression of COX-2 mRNA in U118 cells was significantly down-regulated,and the difference was statistically significant(P<0.05).In western blot experiment,after melatonin combined with nimustine,the expression of COX-2 protein and p-IκBα/IκBα in U118 cells was significantly down-regulated,and the difference was statistically significant(P<0.05).Conclusion Melatonin can enhance the inhibitory effect of nimustine on the proliferation of glioma U118 cells.

关 键 词：胶质瘤 褪黑素 尼莫司汀 环氧化酶-2 核因子ΚB 

分 类 号：R730.264[医药卫生—肿瘤]