CAJ | 학술논문

目的:建立人喉癌长春新碱多药耐药细胞系.方法:以药物浓度梯度递增法诱导筛选人喉癌Hep-2的多药耐药细胞株Hep-2/v,比较两组细胞的形态和倍增时间;MTT法确定细胞的IC50及其耐药指数;流式细胞仪检测两组细胞的细胞周期分布以及细胞内的罗丹明聚集情况;实时定量RT-PCR法检测MDR1基因的mRNA表达,Western blot法检测相应的蛋白表达情况.结果:建成的Hep-2/v耐药细胞株,其耐药性能稳定,耐药指数为45,并与顺铂及5-氟尿嘧啶有不同程度的交叉耐药性;Hep-2/v的体外群体细胞倍增时间较亲代细胞延长13.58小时;细胞周期分析结果显示其G_0/G_1期细胞升高,而S期细胞则明显降低(P＜0.05);罗丹明染色显示,Hep-2细胞内的罗丹明较Hep-2/v明显升高(P＜0.01);Hep-2/v细胞MDR1表达在基因及蛋白水平明显高于Hep-2细胞(P＜0.01).结论:Hep-2/v细胞株具有明确及稳定的多药耐药性,是研究多药耐药机制及筛选逆转剂的良好模型.
목적:건립인후암장춘신감다약내약세포계.방법:이약물농도제도체증법유도사선인후암Hep-2적다약내약세포주Hep-2/v,비교량조세포적형태화배증시간;MTT법학정세포적IC50급기내약지수;류식세포의검측량조세포적세포주기분포이급세포내적라단명취집정황;실시정량RT-PCR법검측MDR1기인적mRNA표체,Western blot법검측상응적단백표체정황.결과:건성적Hep-2/v내약세포주,기내약성능은정,내약지수위45,병여순박급5-불뇨밀정유불동정도적교차내약성;Hep-2/v적체외군체세포배증시간교친대세포연장13.58소시;세포주기분석결과현시기G_0/G_1기세포승고,이S기세포칙명현강저(P＜0.05);라단명염색현시,Hep-2세포내적라단명교Hep-2/v명현승고(P＜0.01);Hep-2/v세포MDR1표체재기인급단백수평명현고우Hep-2세포(P＜0.01).결론:Hep-2/v세포주구유명학급은정적다약내약성,시연구다약내약궤제급사선역전제적량호모형.
Objective:To establish a multidrug resistance cell line from human laryngeal cancer cells by VCR.Methods:Human laryngeal cancer cells(Hep-2) were exposed in stepwise escalating concentration of VCR until the resistant cell(Hep-2/v) line was developed. The IC50 and the resistance folds of multidrug resistance were detected with MTT assay. The differences of cell cycle distribution and Rhodamine accumulation between Hep-2 and Hep-2/v cells were studied through flow cytometry. The MDR1 gene were detected through real-time quantitative RT-PCR, and the corresponding protein was detected through western blot.Results:Hep-2/v cells was established, which had stable resistance to VCR and a resistance index of 45;Hep-2/v cells exhibited cross resistance to many other chemotherapeutic agents and its doubling time was prolonged;The number of cells in G_0/G_1 phase was increased while in S phase decreased(P＜0.05);Rhodamine accumulation in Hep-2 cells was much more than Hep-2/v cells(P＜0.01);The expression of MDR1 were increased than that of Hep-2 cells(P＜0.01).Conclusion:Hep-2/v cell line shows the typical multidrug resistance phenotype.It can be served as a model for the study of MDR.