CAJ | 학술논문

目的建立人类淋巴瘤B JAB多药耐药细胞株BJAB/ADR,并初步研究其耐药机制.方法采用多柔比星浓度梯度递增法建立人类B细胞淋巴瘤耐药细胞模型BJAB/ADR,观察其生长规律并绘制细胞生长曲线;用四甲基偶氮唑蓝(MTT)法鉴定耐药细胞株对多种化疗药物的耐药性并计算耐药指数;提取耐药细胞株RNA,实时定量聚合酶链反应(PCR)法检测相关耐药基因MDR1 mRNA的表达;流式细胞术检测细胞表面P糖蛋白(Pgp)的表达;通过罗丹明外排实验,检测Pgp功能.结果成功建立B细胞淋巴瘤耐药细胞模型BJAB/ADR,并在160 ng/ml多柔比星溶液中稳定生长,耐药细胞较敏感细胞生长缓慢,细胞形态无明显变化.MTT检测结果表明BJAB/ADR细胞对多柔比星的耐药指数为43倍,同时对柔红霉素、依托泊苷、高三尖杉酯碱(HHT)及米托蒽醌(MX)也具有一定的耐药性.实时定量PCR结果表明,耐药细胞BJAB/ADR MDRl mRNA明显高于药物敏感细胞BJAB(P＜ 0.01).流式细胞术检测结果显示,耐药细胞表面Pgp高表达;罗丹明外排实验表明Pgp对多柔比星具有外排功能,使B JAB/ADR细胞获得耐药特性.结论建立了人类B细胞淋巴瘤的耐药细胞株BJAB/ADR,其对多柔比星耐药性稳定,并呈现多药耐药细胞的基本生物学特性,为进一步研究肿瘤多药耐药发生的机制及逆转耐药提供有利的模型.
목적 건립인류림파류B JAB다약내약세포주BJAB/ADR,병초보연구기내약궤제.방법 채용다유비성농도제도체증법건립인류B세포림파류내약세포모형BJAB/ADR,관찰기생장규률병회제세포생장곡선;용사갑기우담서람(MTT)법감정내약세포주대다충화료약물적내약성병계산내약지수;제취내약세포주RNA,실시정량취합매련반응(PCR)법검측상관내약기인MDR1 mRNA적표체;류식세포술검측세포표면P당단백(Pgp)적표체;통과라단명외배실험,검측Pgp공능.결과 성공건립B세포림파류내약세포모형BJAB/ADR,병재160 ng/ml다유비성용액중은정생장,내약세포교민감세포생장완만,세포형태무명현변화.MTT검측결과표명BJAB/ADR세포대다유비성적내약지수위43배,동시대유홍매소、의탁박감、고삼첨삼지감(HHT)급미탁은곤(MX)야구유일정적내약성.실시정량PCR결과표명,내약세포BJAB/ADR MDRl mRNA명현고우약물민감세포BJAB(P＜ 0.01).류식세포술검측결과현시,내약세포표면Pgp고표체;라단명외배실험표명Pgp대다유비성구유외배공능,사B JAB/ADR세포획득내약특성.결론 건립료인류B세포림파류적내약세포주BJAB/ADR,기대다유비성내약성은정,병정현다약내약세포적기본생물학특성,위진일보연구종류다약내약발생적궤제급역전내약제공유리적모형.
Objective To establish human multi-drug resistance (MDR) B-cell lymphoma cell line BJAB/ADR and investigate its biological characteristics.Methods Human B-cell lymphoma MDR cell line BJAB/ADR was induced by exposure to increasing dose of adriamycin.The growing features of BJAB/ADR were observed and cell growth was measured,IC50 of antitumor agents was evaluated by MTT assay.The expression of MDR1 mRNA was determined with real-time PCR,FACS was applied to study the expression of MDR protein P-glycoprotein (Pgp) and Rhodamine 123R efflux assay was used to determine the function of Pgp.Results Human B-cell lymphoma MDR cell line BJAB/ADR was successfully established.MTF results showed BJAB/ADR grew steadily with 160 ng/ml adriamycin in the medium and possessed 45-fold resistance to adriamycin compared with BJAB.Also BJAB/ADR cells exhibited cross-resistance to daunorubicin,etoposide,homohardngtonine and mitoxantrone.In term of the cell proliferation,as compared to BJAB cells,BJAB/ADR cells grew more slowly without obvious changes of cellular morphology.QRT-PCR indicated that the mRNA level of MDR1 in BJAB/ADR resistant cells was significantly higher than that in BJAB cells (P ＜ 0.01).FACS results revealed that the protein expression of Pgp in BJAB/ADR was increased compared with that in BJAB cells.Rhodamine 123R efflux assay indicated Pgp had the capacity of drug excretion and decreased the intracellular drug concentration.Conclusions Human B-cell lymphoma MDR cell line BJAB/ADR is successfully established.BJAB/ADR possesses a stable adriamycin resistance and presents some basic biological characteristics of MDR cells.It offers a model with MDR phenotype for the study of MDR in human B-cell lymphoma.