去甲斑蝥素对人多发性骨髓瘤细胞株U266生长调控的影响及其机制的研究

EFFECT OF NORCANTHARIDIN ON THE GROWTH CONTROL OF HUMAN MYELOMA CELL LINE U266 AND ITS MECHANISM

目的研究去甲斑蝥素(norcantharidin,NCTD)对多发性骨髓瘤细胞株的增殖及其凋亡的影响及其可能的机制。方法四甲基偶氮唑盐[3-(4,5-dimethyl-2 thiahiazoyi)-3,5-di-phenyl-tetrazolium bromide,MTT]检测5、20、40、80、160μmol/L的NCTD对U266细胞增殖的影响;用流式细胞术检测不同浓度NCTD对细胞周期及细胞凋亡的影响,并应用半定量反转录聚合酶链反应检测survivin基因表达的变化。结果在20～160μmol/L的浓度内,NCTD对U266细胞的增殖有抑制,抑制与NCTD的浓度呈剂量依赖关系(P<0.05)。不同浓度的NCTD能诱导多发性骨髓瘤细胞株U266凋亡,使细胞明显阻滞于G_2/M期;在NCTD作用下,U266细胞survivin及其机制可能与诱导凋亡、细胞周期阻滞有关。结论 NCTD能抑制U266细胞的增殖,并诱导其凋亡的机制可能与抑制survivin的表达及G_2/M期阻滞有关。

Objective To study the inhibitory effect of norcantharidin（NCTD） on the growth of human myeloma cell line U266 . Methods U266 cells were treated with different concentrations of NCTD . The growth of U266 cells was measured by MTF（3 , （4,5-dimethyl -2 thiahiazoyi） -3,5 -di - phenyl - tetrazolium bromide ）, and changes of apoptosis rate and cell cycle by flow cytometry. Semiquantitative reverse transcription - polymerase chain reaction was used to detect the effect of NCTD on survivin gene expression. Results Within the concentrations of 20 - 160p, mol/L, NCTD can obviously inhibited the growth of U266 cells in a dose - dependent manner. NCTD can increase Annexin V positive rate, block cell at G2/M phase, reduce the gene expression of survivin . Conclusion NCTD can inhibit the growth of U266 cells in a dose - dependent manner and inhibit survivin gene expression . The mechanism is probably related to apoptosis induction and cell cycle arrest in G2/M phase.