摘要
目的:研究拓扑替康(TPT)对骨髓增生异常综合征(MDS)急性髓系白血病(AML)M2变细胞系MUTZ-1的作用机制,以及环磷酰胺(CTX)对TPT作用靶标Topo-ⅠmRNA表达水平的影响。方法:MTT法测定TPT对MUTZ-1细胞的杀伤作用。流式细胞术分析TPT对MUTZ-1细胞周期的影响。染色体分析方法观察TPT对MUTZ-1的影响。半定量RT-PCR检测CTX对Topo-ⅠmRNA表达水平的影响。结果:①5、10、20、40、80、160nmol/L TPT与MUTZ-1细胞共培养72h后细胞存活率呈剂量依赖性性下降,分别为(78.9±10.7)%、(69.2±4.7)%、(58.6±7.1)%、(49.9±7.6)%、(42.4±0.8)%和(23.7±9.8)%,各实验组与对照组(505.6±20.1)%之间均差异有统计学意义(均P<0.05)。②TPT阻碍MUTZ-1细胞染色体发生分离,并使MUTZ-1细胞发生G2/M期阻滞,该阻滞呈时间依赖性和剂量依赖性。③半定量RT-PCR发现0.224mmol/LCTX与MUTZ-1细胞共培养24、48和72h后,Topo-ⅠmRNA表达都上升,以48h的表达量最高。结论:TPT对MUTZ-1细胞的生长抑制呈剂量依赖性。染色体分离受阻和G2/M期阻滞是TPT的作用机制之一。CTX可上调Topo-Ⅰ表达,与TPT发挥联合抗白血病作用。
Objective：To explore the anti-leukemia effect of TPT on MUTZ-1 cells（MDS-RAEB transforming to AML cell line）in vitro,and the effect of cyclophosophamide（CTX）on the level of Topo-ⅠmRNA in MUTZ-1 cells.Method：The cytotoxic effect of TPT on MUTZ-1 cells was determined by MTT assay.The changes of cell cycle were observed by FCM with PI staining.The karyotypic changes of TPT on MUTZ-1 cells were observed by karyotyping.The effects of CTX on the level of Topo-ⅠmRNA were determined by hemi-quantitative RT-PCR.Result：①After incubation of MUTZ-1 cell with 5,10,20,40,80 and 160nmol/L TPT for 72h,the survival ratesprogressively reduced to（78.9±10.7）%,（69.2±4.7）%,（58.6±7.1）%,（49.9±7.6）%,（42.4±0.8）% and（23.7±9.8）%（P〈0.05）,compared with control（505.6±20.1）%.② FCM with PI staining showed that more MUTZ-1 cells were arrested in G2/M phase,which was time-dependent and dose-dependent.Karyotyping results showed blockage of chromosome segregations.③As revealed by hemi-quantitative RT-PCR there was an increasing trend in the level of Topo-ⅠmRNA in MUTZ-1 treated with 0.224 mmol/L CTX for 24 h,48 h and 72 h.Conclusion：The inhibiting effect of TPT on MUTZ-1 activity is dose-dependent.TPT induces blockage of chromosome segregations and G2/M phase arrest in MUTZ-1 cells.CTX can upregulate the level of Topo-ⅠmRNA,which has synergism with TPT.
出处
《临床血液学杂志》
CAS
2012年第4期457-460,共4页
Journal of Clinical Hematology
