环氧化物酶抑制剂NS-398联合奥沙利铂对宫颈癌HeLa细胞生长抑制及诱导凋亡作用的研究  

Effect of cyclooxygenase inhibitor NS-398 combined with Oxaliplatin on growth inhibition and apoptosis induction of HeLa cells in cervical cancer

作　　者：王静[1] 金燕 董尚林 武欣[4] WANG Jing;JIN Yan;DONG Shanglin;WU Xin(Department of Gynaecology and Obstetrics,the First Affiliated Hospital of Hebei North University,Heibei Province,Zhangjiakou 075000,China;Life Science Research Center,Hebei North University,Heibei Province,Zhangjiakou 075000,China;the First Affiliated Hospital of Hebei North University,Heibei Province,Zhangjiakou 075000,China;Department of Pathology,Hebei North University,Heibei Province,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院附属第一医院妇产科,河北张家口075000 [2]河北北方学院生命科学研究中心,河北张家口075000 [3]河北北方学院附属第一医院,河北张家口075000 [4]河北北方学院病理教研室,河北张家口075000

出　　处：《中国医药导报》2021年第3期30-34,F0004,共6页China Medical Herald

基　　金：河北省卫生健康委员会医学科学研究课题计划项目(20190870)。

摘　　要：目的研究环氧化酶抑制剂NS-398联用奥沙利铂(L-OHP)对宫颈癌HeLa细胞的抑制及诱导凋亡作用。方法体外培养人宫颈癌HeLa细胞,CCK-8法检测不同浓度NS-398(0、30、60μmol/L),L-OHP(0.0、2.5、5.0、10.0 mg/L)单独作用及联合作用于HeLa细胞后对细胞的抑制作用;将HeLa细胞随机分为:对照组、NS-398组(60μmol/L)、L-OHP组(10 mg/L)及联合组(NS-39860μmol/L+L-OHP 10 mg/L),倒置显微镜观察HeLa细胞形态改变,罗丹明染色观察HeLa细胞凋亡情况,流式细胞仪检测HeLa细胞早期凋亡率,DNA琼脂糖凝胶电泳检测梯状条带。结果CCK-8结果显示,与未加药物组比较,NS-398单药组和L-OHP单药组抑制率增高;联合组抑制率高于NS-398单药组和L-OHP单药组(均P<0.01)。对照组细胞状态良好,形态呈梭形,贴壁生长,细胞边缘清晰,胞膜完整。NS-398组细胞体积皱缩变小变圆,胞质内出现颗粒状物质,少量细胞从瓶底脱落,漂浮于培养液中。L-OHP组细胞失去正常形态,皱缩变小,培养液中可见死细胞。联合组培养液中可见大量死细胞和细胞碎片,仅余少量细胞贴在瓶底,且形态呈不同程度的皱缩,部分细胞破裂,细胞内容物溢出。罗丹明123染色结果显示,对照组无荧光;NS-398组、L-OHP组与联合组荧光强度增强,尤其以联合组最为明显。流式细胞仪检测结果显示,NS-398组和L-OHP组凋亡率明显高于对照组;联合组凋亡率明显高于NS-398组、L-OHP组(均P<0.01)。DNA电泳显示,联合组出现了明显的梯状条带,NS-398组和L-OHP组有明显的弥散现象出现;对照组即无弥散现象也无梯状条带出现。结论NS-398能够增强L-OHP对HeLa细胞的生长抑制作用和诱导凋亡作用。Objective To explore the effect of cyclooxygenase inhibitor NS-398 combined with Oxaliplatin(L-OHP)on growth inhibition and apoptosis induction of HeLa cells in cervical cancer.Methods HeLa cells were cultured in vitro.CCK-8 method was used to detect the inhibitory effect of different concentrations of NS-398(0,30,60μmol/L)and L-OHP(0.0,2.5,5.0,10.0 mg/L)on HeLa cells alone or in combination.HeLa cells were divided into:control group,NS-398 group(60μmol/L),L-OHP group(10 mg/L)and combined group(NS-39860μmol/L+L-OHP 10 mg/L).The morphological changes of HeLa cells were observed by inverted microscope.The apoptosis of HeLa cells was observed by Rhodamine staining,and the early apoptosis rate of HeLa cells was detected by flow cytometry.The ladder strip was detected by DNA agarose gel electrophoresis.Results CCK-8 analysis showed that compared with untreated group,the cell growth inhibition rate in NS-398 single drug group and L-OHP single drug group were higher;growth inhibition rate in combined group was higher than that in NS-398 single drug group and L-OHP single drug group(all P<0.01).The cells in control group were in good condition,with fusiform shape,adherent growth,clear cell edges and intact cell membrane.In NS-398 group,the volume of cells shrank and became round,granular substances appeared in the cytoplasm,and a small number of cells fell off from the bottom of the bottle and floated in the culture medium.The cells of L-OHP group lost their normal morphology and shrank,and dead cells could be seen in the culture medium.A large number of dead cells and cell debris could be seen in the culture medium of the combined group,with only a few cells sticking to the bottom of the bottle,with different degrees of shrinkage,partial cell rupture,and cell contents overflow.Rodamine 123 staining result showed that,the control group had no fluorescence.The fluorescence intensity of NS-398 group,L-OHP group and combination group was enhanced,especially the combination group.The results of flow cytometry showed that

关 键 词：NS-398 奥沙利铂 宫颈癌细胞 凋亡 

分 类 号：R914[医药卫生—药物化学]