摘要
目的探讨阿糖胞苷(Ara-C)协同地塞米松(DEX)诱导糖皮质激素(GC)抵抗人类T淋巴细胞ALL细胞凋亡的效应及其临床意义。方法将不同水平的Ara-C(30 nmol.L-1、100 nmol.L-1、300 nmol.L-1)分别与不同浓度的DEX(100μmol.L-1、500μmol.L-1)联用对GC抵抗型Jurkat细胞进行不同浓度联合药物暴露。通过流式细胞计数检测各组细胞24 h、48 h暴露后凋亡率;ELISA法检测暴露24 h后核因子-κB(NF-κB)p65亚基活性;反转录-PCR(RT-PCR)和Western blot法检测暴露24 h后Survivin表达和蛋白水平。结果 Ara-C(100 nmol.L-1、300 nmol.L-1)与DEX(100μmol.L-1、500μmol.L-1)联合暴露组,其24 h和48 h暴露后细胞凋亡率均显著高于同浓度DEX单药暴露组(Pa<0.001),且呈显著浓度依耐性;联合暴露组NF-κB p65活性和Sur-vivin转录水平及蛋白水平较同浓度DEX单药暴露组均有显著下降(Pa<0.05)。结论 Ara-C与DEX具有显著的浓度依赖协同诱导GC抵抗型Jurkat细胞凋亡的作用,且这种协同效应可能是由于二者对NF-κB活性及其下游Survivin表达的协同抑制所产生的。
Objective To explore the synergistic effect of cytosine arabinoside（Ara-C） combined with dexamethasone（DEX） on inducing glucocorticoid against the apoptosis of human T-acute lymphoblastic leukemia cells and its clinical significance.Methods The Jurkat cells were exposed to DEX（100 μmol·L-1,500 μmol·L-1） combination with Ara-C（30 nmol·L-1,100 nmol·L-1,300 nmol·L-1）.The cell apoptosis was assayed by flow cytometry after 24 h and 48 h exposure;the nuclear factor（NF）-κB p65 subunit viability was detected by enzyme-linked immunosorbent assay after 24 h exposure;the Survivin gene expression and the protein levels were detected by reverse transcription-polymerase chain reaction and Western blot after 24 h exposure.Results After 24 h and 48 h exposure in Ara-C（100 nmol·L-1,300 nmol·L-1） and DEX（100 μmol·L-1,500 μmol·L-1） combined exposed subgroups,the cell apoptosis rate was significantly higher than the same concentration of DEX single-exposed subgroup（Pa0.001）,and the synergistic effect was concentration dependent;and the NF-κB p65 viability,survivin transcription and protein levels were also significantly decreased too after 24 h combined exposure（Pa0.05）.Conclusions The Ara-C combined with DEX can significantly synergistically induce the glucocorticoid-resisted Jurkat cells apoptosis,and this effect is concentration dependent;this effect may be associated with the synergistic inhibition of the NF-κB viability and Survivin gene expression which may download of NF-κB.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第3期166-168,共3页
Journal of Applied Clinical Pediatrics
基金
湖北省卫生厅科研重点项目(JX4A05)
