摘要
目的研究氟达拉宾（fludarabine，FDB）联合格列卫（ST1571）诱导对慢性粒细胞白血病（CML）细胞株K562耐药性的影响。方法通过逐步增加STI571的浓度诱导培养耐药性K562-R细胞，在STl571存在的情况下，用不同浓度（0，1，2，3，4，5μmol／L）的FDB药物处理K562-R细胞，作用24h后，收集细胞，进行Hoechst细胞染色计数，流式细胞仪检测肿瘤多药耐药基因MDR-1的表达，实时定量PCR（RT—PCR）检测bcr-abl融合基因，细胞形态观察，westernblot检测相关凋亡蛋白的表达。结果成功得到K562-R细胞株，该细胞可以在浓度为0．8μmol／L的ST1571中稳定生长并且可以快速增殖，经过FDB药物处理K562-R细胞24h后发现，实验中任何浓度的FDB均可降低K562-R细胞的耐药性，抑制细胞的生长，抑制率分别为23％，36％，47％，78％和93％，抑制率与FDB浓度呈正相关。并促使细胞进入凋亡期，相关凋亡蛋白的表达均能检测到，同时可以降低MDR-1和ber-abl融合基因的表达，细胞形态观察也显示细胞出现大量死亡，K562-R的耐药性明显降低。结论FDB联合ST1571诱导K562细胞，可以明显降低K562细胞的耐药性，提高治疗效果，可为临床抗肿瘤免疫治疗提供新的借鉴与思路。
Objective In order to research the effects of chronic myeloid leukemia （CML） K562 cell lines resistance induced by FDB joint STI571. Methods Inductied of resistance K562-R cells by gradually increasing the concentration of STI571, FDB drug treatment with different concentrations （0, 1,2,3,4,5 μmol/L） of K562-R cells for 24 h in the presence of STI571 ,then collected the cells,dyeing counted the cells by Hoechst. Detected the expression of MDR-1 by flow cytometry and the ber-abl fusion gene by real-time quantitative PCR （RT-PCR）. Observed the cell morphology and detected the expres- sion of apoptotic protein through western blot. Results Finally,got the KS62-R cell lines,the cells in stable growth and rap- id proliferation at the concentration of 0.8 t^mol/L STI571, after the FDB drug treatment of K562-R cells for 24 h, found that any concentration of the FDB could reduce the resistance o5 the K562-R cells to STI571, the inhibition rates were 23 ~, 36 ~//0,47 ~/0,78% and 93 %, respectively. The inhibition rate was positively correlated with FDB concentration, and promote cells to apoptosis,related apoptosis protein expression also could be detected, while the expression of MDR-1 and bcr-abl fu- sion gene decreased. The resistance of K562-R significantly reduced and cell apoptosis appeared. Conclusion Using the FDB Joint STI571 in the induction of K562 cells,can significantly reduce the resistance of K562 cells and improve the therapeutic effect. This method can be effective as a clinical anti-tumor immune treatment.
出处
《现代检验医学杂志》
CAS
2012年第5期11-14,共4页
Journal of Modern Laboratory Medicine
基金
陕西省科技厅社发公关项目（2012KB-02-29）.
