CD44对K562细胞株耐药的影响及其机制

发布时间：2018-04-08 17:38

本文选题：白血病　切入点：耐药　出处：《延边大学》2017年硕士论文

【摘要】：目的:探讨CD44在K562细胞及阿霉素耐药细胞株K562/ADR中的表达情况,阐明CD44与K562细胞耐药之间的相关性,探索逆转白血病耐药的途径。方法:1.利用实时荧光定量PCR法,检测K562细胞及K562/ADR细胞中CD44的mRNA表达水平;2.用电转染法,制备高表达CD44的K562细胞(K562/CD44细胞);3.用倒置荧光显微镜下观察法、实时荧光定量PCR法,检测K562和K562/CD44细胞的CD44标记绿色荧光蛋白、CD44的mRNA表达水平,确认转染成功与否;4.WST-1细胞增殖实验,检测K562细胞的阿霉素IC50,选择适宜的阿霉素药物浓度和作用时间,并对比K562和K562/CD44细胞在阿霉素相同浓度和相同时间作用下的存活率。结果:1.实时荧光定量PCR检测结果:与K562细胞比较K562/ADR细胞中的CD44 mRNA表达水平显著上升(P0.01);2.倒置荧光显微镜观察结果显示:CD44高表达载体转染后的细胞具有绿色荧光蛋白的高表达;3.实时荧光定量PCR检测结果:与K562细胞相比较K562/CD44细胞中的CD44 mRNA表达显著上调(P0.01);4.WST-1 细胞增殖实验检测结果:(1)0.25mg/L、0.5mg/L、1.Omg/L、1.5mg/L和2.0mg/L的阿霉素作用于K562细胞24h、48h、72h均显著抑制细胞增殖,并呈时间和浓度依赖性,1.0mg/L、1.5mg/L和2.0mg/L的阿霉素抑制细胞增殖效应无显著差异(P0.05),K562细胞的阿霉素IC50为0.7376mg/L;(2)与K562细胞比较,K562/CD44细胞在1.0mg/L阿霉素作用48h时的存活率显著上升(P0.01)。结论:1.CD44过表达的K562细胞对阿霉素的敏感性减弱;2.CD44的过表达可能参与了 K562细胞的耐药机制。
[Abstract]:Objective: to investigate the expression of CD44 in K562 cells and doxorubicin resistant cell line K562/ADR, to elucidate the correlation between CD44 and drug resistance of K562 cells, and to explore the way to reverse the drug resistance of leukemia.Method 1: 1.The mRNA expression of CD44 in K562 cells and K562/ADR cells was detected by real-time fluorescence quantitative PCR assay.K562 cells with high expression of CD44 were prepared by electrotransfection.The expression of CD44 labeled green fluorescent protein CD44 (CD44) in K562 and K562/CD44 cells was detected by reverse fluorescence microscope and real-time fluorescence quantitative PCR, and the proliferation of K562 and K562/CD44 cells was confirmed by 4.WST-1 cell proliferation.The doxorubicin IC50 of K562 cells was detected, and the optimal concentration and time of doxorubicin treatment were selected, and the survival rate of K562 and K562/CD44 cells was compared under the same concentration and time of doxorubicin.The result is 1: 1.Results of real-time fluorescence quantitative PCR: compared with K562 cells, the expression of CD44 mRNA in K562/ADR cells increased significantly (P 0.01).The results of inverted fluorescence microscope showed that the cells transfected with the high expression vector of 1% CD44 had high expression of green fluorescent protein (GFP).Conclusion the over-expression of CD44 in K562 cells may be involved in the mechanism of drug resistance in K562 cells.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.7

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