摘要
目的探讨Survivin基因在白血病糖皮质激素(GC)敏感型(CEM)和GC耐药型(Jurkat)细胞株中的表达差异及其在GC耐药中的临床意义。方法实验分为未暴露组和暴露组,并设阴性对照及空白对照。应用四甲基偶氮唑蓝(MTT)法检测细胞增殖情况,流式细胞术分析细胞凋亡率,反转录PCR(RT-PCR)检测Survivin基因mRNA水平,Western blot法分析蛋白水平。结果 1.MTT结果显示:地塞米松磷酸钠(DEX)以剂量、时间依赖方式抑制CEM和Jurkat细胞株增殖。CEM和Jurkat的24h抑制细胞增殖50%的药物浓度(IC50)分别为442μmol.L-1和13μmol.L-1,其差异有统计学意义(P<0.01);CEM和Jurkat的48hIC50分别为273μmol.L-1和3μmol.L-1,其差异有统计学意义(P<0.01)。DEX对Jurkat的抑制增殖效应远较CEM差。2.流式细胞仪分析结果证实:DEX对Jurkat的诱导凋亡作用显著低于CEM(P<0.01)。3.mRNA和蛋白水平结果均显示:未暴露组Jurkat细胞株Sur-vivin表达水平较CEM显著升高(P<0.05)。暴露组CEM细胞株Survivin表达变化较Jurkat细胞株显著(P<0.01)。结论 Sur-vivin基因在GC敏感型和GC耐药型白血病细胞株中的表达差异可能与细胞株对GC的敏感性相关。其在GC耐药型白血病细胞株中的持续高表达可能是GC耐药发生的机制之一。
Objective To investigate the different expressions of Survivin gene in glucocorticoid（GC）-sensitive and GC-resistant cell line and their clinical significance.Methods Samples were divided into unexposed group and exposed group,and also the negative control and the blank controls were established.Inhibition of cell proliferation was assayed by methyl thiazolyl tetrazolium（MTT）;Cell apoptosis was examined by flow cytometry;Survivin mRNA level was detected by RT-PCR;Survivin protein was evaluated by Western blot.Results 1.The inhibition effect of dexamethasone sodium phosphate（DEX） was dose-time dependent,50% drug concentration of Jurkat and CEM′s 24 h inhibitory cells proliferation （IC50） was 442 μmol·L^-1 and 13 μmol·L^-1（P〈0.01）;Jurkat and CEM′s 48 h IC50 was 273 μmol·L^-1and 3 μmol·L^-1（P〈0.01）.The effect of DEX on Jurkat proliferation induction was lower than that of GEM.2.Flow cytometry showed the apoptosis rate of CEM treated with DEX was significantly higher than that of Jurkat（P〈0.01）.3.The level of Survivin expression in Jurkat cell was significantly higher than that of CEM in unexposed group（P〈0.05）;Survivin expression in CEM cell was significantly decreased in exposed group （P〈0.01）.Conclusions The different levels of Survivin gene in GC-sensitive and GC-resistant leukemia cell line might be reflected to the GC sensitivity of leukemia cells.Survivin overexpression persisting in leukemia cells may lead to GC resistance.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2010年第15期1136-1139,共4页
Journal of Applied Clinical Pediatrics
基金
湖北省卫生厅科研重点项目(JX4A05)
