辛伐他汀对急性早幼粒细胞白血病NB4细胞NF-κB信号通路的影响

Effects of simvastatin on NF-κB signaling pathways in human acute promyelocytic leukemia NB4 cell line

目的探讨辛伐他汀对人急性早幼粒细胞白血病细胞株(NB4细胞)增殖与凋亡的影响以及对凋亡信号转导通路NF-κB通路相关基因mRNA表达的影响,以探讨辛伐他汀抗白血病效应及可能机制。方法以不同浓度辛伐他汀处理NB4细胞,取对数生长期细胞进行实验。实验分为阴性对照组和辛伐他汀处理组(终浓度分别为5、10、15μmol/L),培养24、48、72 h。采用MTT法观察NB4细胞增殖能力;流式细胞术测定NB4细胞凋亡指标Annex-inⅤ/propidium iodide变化;PCR芯片研究NB4细胞NF-κB通路相关基因mRNA的差异表达。结果辛伐他汀对NB4细胞的生长有明显抑制作用和促凋亡作用(均P<0.01),且呈时间与剂量依赖性。15μmol/L辛伐他汀处理NB4细胞24、48、72 h细胞抑制率分别达到45.75%、92.91%、96.46%,而细胞早期凋亡率分别为30.25%、64.34%、87.38%。与对照组相比,15μmol/L辛伐他汀处理NB4细胞48 h组中NF-κB通路有56个基因表达发生改变,其中47个基因表达下调、9个基因表达上调。结论辛伐他汀能抑制NB4细胞增殖并诱导其凋亡,其诱导凋亡机制可能与辛伐他汀调节NF-κB通路相关基因的表达有关。

Objective To investigate the effects of simvatatin on proliferation, apoptosis, the NF-KB signaling pathway in human acute promyelocytic leukemia cell lines NB4. Methods NB4 cells were incubated with different concentration of simvastatin （5,10,15 μmoL/L）, NB4 cells without any treatment were as control. Cells of different detection. MTF method was used groups were collected at 24, 48 to evaluate the growth inhibition and 72 h after incubation for further rate and flow cytometry was used to detect the early stage apoptosis and necrosis ratio. The human NF-KB signaling array profiles the expression of genes involved in NF-s：B signaling. Results pathway RT2 profiler PCR Simvastatin inhibited the proliferation and induced the apoptosis of NB4 cells in time and dose-dependent manner significantly （ all P 〈 0. 01 ）. The cell growth inhibition rate of NB4 cells treated with 15μmol/L simvastatin were 45.75%, 92.91% and 96.46% respectively at 24, 48 and 72 h; and the early stage apoptosis ratio were 30.25%, 64.34% and 87.38% , respectively. Compared with the control group, 56 genes expression levels changed in the group of 15 μmol/L simvatatin group at 48 h, among which, 47 genes were down- regulated and 9 genes were up-regulated. Conclusion Simvatatin potentially inhibited proliferation and induce apoptosis of NB4 cells, and the mechanism may be associated with the changes of gene expression levels involved in NF-KB signaling pathway regulated by simvastatin.