双氢青蒿素对人口腔鳞癌细胞KBV200多药耐药的影响  

Effect of dihydroartemisinin on multidrug resistance of human oral squamous cell carcinoma cell line KBV200 by regulating ROS-MAPK pathway

作　　者：刘族志 赵永兴 林建能 周国平 LIU Zu-zhi;ZHAO Yong-xing;LIN Jian-neng;ZHOU Guo-ping(Department of Stomatology,Danzhou People's Hospital,Danzhou 571700,Hainan Province,China)

机构地区：[1]儋州市人民医院口腔科

出　　处：《上海口腔医学》2019年第6期586-590,共5页Shanghai Journal of Stomatology

基　　金：海南省卫生计生行业科研项目（16A200063）

摘　　要：目的:检测双氢青蒿素(dihydroartemisinin,DHA)对人口腔鳞癌细胞KBV200多药耐药的作用,并探讨其与ROS-MAPK通路的关系。方法:利用MTT法检测不同浓度DHA对KB、KBV200细胞增殖的影响,顺铂(cisplatin,DDP)、长春新碱(vincristine,VCR)、阿霉素(adriamycin,ADM)、依托泊苷(etoposide,VP-16)对KB、KBV200细胞的增殖抑制率及加入DHA后的变化;分别联合ROS诱导剂3-AT、ERK抑制剂UO126、JNK抑制剂SP600125、p38MAPK抑制剂SB203580,观察干预前、后的逆转差异。利用流式细胞术检测各组细胞内ROS的荧光强度,采用SPSS 17.0软件包对数据进行统计学分析。结果:与KB细胞相比,KBV200细胞对VCR、VP-16、ADM药物的耐药性显著提高(P<0.05),10、20、30μg/mL DHA作用后,KBV200细胞对VCR、VP-16、ADM的IC50显著降低,呈浓度依赖性(P<0.05)。与KB组细胞相比,KBV200组细胞ROS荧光强度显著降低(P<0.05);DHA处理后,ROS荧光强度显著增高,VCR、VP-16、ADM IC50显著降低(P<0.05),与ROS促进剂效果一致。与KB细胞相比,KBV200细胞p-ERK/ERK、p-JNK/JNK、p-p38MAPK/p38MAPK水平显著升高(P<0.05);DHA处理后,p-ERK/ERK、p-JNK/JNK、p-p38MAPK/p38MAPK水平显著降低,VCR、VP-16、ADM IC50显著增高(P<0.05);加入ERK抑制剂UO126、JNK抑制剂SP600125、p38MAPK抑制剂SB203580后,KBV200细胞VCR、VP-16、ADM IC50进一步降低。结论:双氢青蒿素可能通过促进ROS生成、抑制MAPK通路而逆转KBV2001细胞的多药耐药性。PURROSE:To investigate the effect of dihydroartemisinin(DHA)on multidrug resistance of human oral squamous cell carcinoma cell line KBV200 and to explore its relationship with ROS-MAPK pathway.METHODS:MTT assay was used to detect the effects of different concentrations of DHA on proliferation of KB and KBV200 cells.MTT assay was used to detect the inhibition rates of cisplatin(DDP),vincristine(VCR),adriamycin(ADM),etoposide(VP-16)on proliferations of KB and KBV200 cells and the changes after DHA addition,combined with ROS inducer 3-AT,ERK inhibitor UO126,JNK inhibitor SP600125,and p38MAPK inhibitor SB203580,respectively,the difference of reversal before and after intervention was observed;the fluorescence intensity of ROS was measured by flow cytometry.SPSS 17.0 software package was used to analyze the data.RESULTS:Compared with KB cells,drug resistances of KBV200 cells to VCR,VP-16 and ADM were significantly increased(P<0.05),after 10,20,and 30μg/mL DHA treatment;the IC50 of KBV200 cells to VCR,VP-16 and ADM was decreased significantly in a concentration-dependent manner(P<0.05).Compared with KB cells,ROS fluorescence intensity of KBV200 cells decreased(P<0.05);after DHA treatment,ROS fluorescence intensity increased significantly,IC50 of VCR,VP-16,ADM decreased significantly(P<0.05),which was consistent with ROS promoter effect.Compared with KB cells,the levels of p-ERK/ERK,p-JNK/JNK,p-p38MAPK/p38MAPK in KBV200 cells increased significantly(P<0.05);after DHA treatment,the levels of p-ERK/ERK,p-JNK/JNK,p-p38MAPK/p38MAPK decreased significantly,IC50 of VCR,VP-16,and ADM increased significantly(P<0.05);after adding ERK inhibitor UO126,JNK inhibitor SP600125,and p38MAPK inhibitor SB203580,IC50 of VCR,VP-16,and ADM to KBV200 cells were further reduced.CONCLUSIONS:Dihydroartemisinin may reverse multidrug resistance of KBV2001 cells by promoting ROS production and inhibiting MAPK pathway.

关 键 词：口腔鳞癌 双氢青蒿素 ROS MAPK通路 多药耐药 

分 类 号：R739.8[医药卫生—肿瘤]