AEG-1在5-氟尿嘧啶诱导的人口腔鳞状细胞癌细胞耐药中的作用  

Role of AEG-1 in 5-fluorouracil resistance in human oral squamous cell carcinoma cells

作　　者：刘亭亭 王婷[1] 贾冬萍 王秀梅[1] Liu Tingting;Wang Ting;Jia Dongping;Wang Xiumei(Department of Dentistry,the Second Affiliated Hospital of Harbin Medical University,Heilongjiang Harbin 150086,China)

机构地区：[1]哈尔滨医科大学附属第二医院牙体牙髓科,黑龙江哈尔滨150086

出　　处：《现代肿瘤医学》2020年第12期1995-1999,共5页Journal of Modern Oncology

基　　金：黑龙江省自然科学基金重点项目(编号:ZD2019H002);哈尔滨市科技局科技创新人才资助项目(编号:2016RAXYJ094)。

摘　　要：目的:探讨星形胶质细胞升高基因-1(astrocyte elevated gene-1,AEG-1)对5-氟尿嘧啶(5-FU)诱导的口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞系耐药性的影响。方法:验证高(SCC15)、中(转染空质粒的对照组SCC15、CAL27)、低(CAL27)四组不同AEG-1表达量细胞模型AEG-1基因转染效果。MTT实验观察四组细胞对5-FU耐受性的变化;蛋白质印迹法和细胞划痕实验观察四组细胞在5-FU作用下caspase-3、MMP-2、MMP-9蛋白表达水平变化及细胞迁移能力的变化。结果:AEG-1基因在细胞中转染成功。MTT实验显示不同浓度5-FU作用四组细胞后,过表达组细胞的半数抑制浓度(IC50)为423.7μg/ml,其对照组为213.5μg/ml(P<0.05);沉默细胞组IC50值为291.6μg/ml,其对照组为463.1μg/ml(P<0.05)。Western blot显示过表达组及其对照组中5-FU的浓度分别为400μg/ml和200μg/ml时,MMP-2、MMP-9及caspase-3的表达水平开始升高(P<0.05);沉默组及其对照组中5-FU的浓度分别为240μg/ml和480μg/ml时,MMP-2、MMP-9及caspase-3的表达水平开始升高(P<0.05)。划痕实验显示加入相同浓度5-FU后过表达组细胞迁移率明显高于其对照组(P<0.05);而沉默组细胞迁移率明显低于其对照组(P<0.05)。结论:上调/下调AEG-1表达能增加/降低OSCC细胞对5-FU的耐药性,提示OSCC细胞的耐药性可能与AEG-1表达量相关,OSCC细胞系中AEG-1的表达情况可能能间接干扰临床药物化疗的效果。Objective:To investigate the effect of AEG-1 on 5-fluorouracil(5-FU)-induced oral squamous cell carcinoma cell line resistance.Methods:Establish a high(SCC15),medium(control group SCC15,CAL27 transfected empty plasmid),low(CAL27)4 groups of different AEG-1 expression cell models,verify the transfection efficiency of AEG-1 gene.MTT assay was used to observe the changes of tolerance to 5-FU in the 4 groups of cells.Western blot and cell scratching experiments were performed to observe the changes of caspase-3,MMP-2 and MMP-9 protein expression levels and cell migration ability in groups of 4 cells.Results:The AEG-1 gene was successfully transfected in cells.MTT assay showed that the concentration of half-inhibition rate(IC50)of the over-expressing group was 423.7μg/ml,and that of the control group was 213.5μg/ml(P<0.05).The IC50 value of the silenced cell group was observed.The ratio was 291.6μg/ml,and the control group was 463.1μg/ml(P<0.05).Western blot showed that the expression levels of MMP-2,MMP-9 and caspase-3 began to increase when the concentration of 5-FU in the overexpression group and the control group was 400μg/ml and 200μg/ml,respectively(P<0.05).The expression levels of MMP-2,MMP-9 and caspase-3 began to increase when the concentration of 5-FU was 240μg/ml and 480μg/ml in the silencing group and the control group,respectively(P<0.05).The scratch test showed that the cell migration rate of the overexpressing group was significantly higher than that of the control group(P<0.05),and the cell migration rate of the silencing group was significantly lower than that of the control group(P<0.05).Conclusion:Up-regulation/down-regulation of AEG-1 expression can increase/reduce the resistance of OSCC cells to 5-FU,suggesting that the resistance of OSCC cells may be related to AEG.The expression level of AEG-1 is related,and the expression of AEG-1 in OSCC may indirectly interfere with the effect of clinical drug chemotherapy.

关 键 词：星形细胞上调基因-1 耐药性 5-氟尿嘧啶 细胞凋亡 

分 类 号：R739.85[医药卫生—肿瘤]