摘要
目的 研究地塞米松 (DXM)和长春新碱 (VCR)对高三尖杉酯碱 (HH)诱导白血病细胞凋亡与核因子 κB (NF κB)活化的影响。方法 采用TdT介导的dUTP缺口末端标记技术(TUNEL)、DNA电泳方法观察HH诱导K5 6 2 n细胞凋亡 ,采用电泳迁移率变动分析 (EMSA)观察HH诱导K5 6 2 n细胞NF κB活化。结果 用 (0 5、5、5 0 ) μmol/L的HH均能诱导K5 6 2 n细胞凋亡率分别为 (30 0 0± 3 34,4 7 13± 3 18,6 8 6 3± 8 14 ) % ,与对照组相比 ,有良好的浓度依赖关系 (P <0 0 5 ) ;DXM 1μmol/L和VCR 0 1μmol/L本身无诱导K5 6 2 n细胞凋亡的作用 ,但均能增强HH 0 5μmol/L诱导的K5 6 2 n细胞凋亡 ,凋亡增加率分别为 85 8%和 114 6 % (P值均 <0 0 5 )。K5 6 2 n细胞未经药物诱导NF κB也有轻度活化 ;HH 0 5 μmol/L可明显诱导K5 6 2 n细胞NF κB活化 ,DXM 1μmol/L和VCR 0 1μmol/L能显著抑制HH 0 5 μmol/L诱导的NF κB活化 ,抑制率分别为 32 0 %和39 4 % (P值均 <0 0 5 )。结论 HH诱导K5 6 2 n细胞凋亡的同时激活NF κB ;DXM和VCR可通过抑制NF κB活化 ,增强其诱导K5 6 2 n细胞凋亡的作用。
Objective To explore the effects of dexamethasone(DXM) and vincristine(VCR) on apoptosis of K562-n cells and activation of nuclear factor-κB-gene binding(NF-κB)in K562-n cells induced by homoharringtonine. Methods K562-n cells were cultured in RPMI-1640 medium. Homoharringtonine of various concentrations was added into the cultures. Twelve hours later, the apoptosis induced by homoharringtonine in K562-n cells was analysed by TdT-mediated X-dUTP nick and end labeling (TUNEL) and DNA electrophoresis. Another sample of K562-n cells was culture together with DXM (1 μmol/L) or VCR (0.1 μmol/L) for 2 hours, then homoharringtonine was added into the cultures. Twelve hours later, the apoptosis in K562-n cells was analysed by TUNEL and DNA electrophoresis. Still another sample of K562-n cells was cultured for 3 hours with homoharringtonine, then electrophoretic mobility shift assay (EMSA) was conducted to determine the DNA-binding activation of NF-κB. A fourth sample of K562-n cells was cultured together with DXM (1 μmol/L) or VCR (0.1 μmol/L) for 2 hours, then homoharringtonine was added into the cultures. Three hours later, EMSA was conducted. Results The apoptosis rates of K562-n cells induced by homoharringtonine of various concentrations were (30.00±3.34)%, (47.13±3.18)% and(68.63±8.14)%, respectively. The apoptosis rates of K562-n cells induced by homoharringtonine being pre-processed with DXM or VCR were (55.75±3.88)% and (64.38±4.60)%, respectively,being 85.8% and 114.6% higher than that induced by homoharringtonine alone (30.00±3.34) % ( all P <0.05). Activation of NF-κB in K562-n cells was induced significantly by homoharringtonine. Activation of NF-κB in K562-n cells induced by homoharringtonine could be suppressed by being pre-processed with 1.0 μmol/L DXM or 0.1 μmol/L VCR. The rate of suppression was 32.0% and 39.4% respectively. Conclusion Homoharringtonine induces apoptosis of K562-n cells and induces NF-κB activation in K562-n cells. The mechanism of increased apoptosis of K562-n cells with DXM or VCR may be related to suppression of activation of NF-κB of K562-n cells.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2003年第5期292-295,共4页
Chinese Journal of Internal Medicine
基金
国家自然科学基金资助项目 (3 9770 3 3 0 )
