RSK抑制剂BI-D1870对急性白血病细胞增殖的影响

Effect of RSK inhibitor BI-D1870 on proliferation of acute leukemia cells

目的 :探讨p90核糖体S6激酶(p90 ribosomal S6 kinase,RSK)抑制剂BI-D1870对急性白血病(acute leukemia,AL)细胞增殖、细胞凋亡和细胞周期分布的影响。方法:应用实时荧光定量PCR法检测6种AL细胞(Molt-4、Jurkat、Nalm-6、THP1、U937和NB-4)中RSK1和RSK2 m RNA的表达水平,CCK-8法检测RSK抑制剂BI-D1870对AL细胞增殖的影响以及BI-D1870、依托泊苷单药和BI-D1870联合依托泊苷对Molt-4细胞的增殖抑制作用。应用FCM法检测BI-D1870对Molt-4细胞凋亡及细胞周期分布的影响以及BI-D1870、依托泊苷单药和BI-D1870联合依托泊苷对Molt-4细胞凋亡的影响。结果:Molt-4和THP1细胞中RSK1 m RNA的表达水平以及Molt-4、Jurkat、THP1、U937和NB-4细胞中RSK2 m RNA的表达水平均高于健康志愿者外周血单核细胞(P值均<0.05)。RSK抑制剂BI-D1870可抑制Molt-4、Jurkat、Nalm-6、THP1、U937和NB-4细胞的增殖,其抑制作用随着作用时间的延长而递增(P<0.05)。BI-D1870联合依托泊苷处理组Molt-4细胞的增殖抑制率明显高于BI-D1870和依托泊苷单药处理组(P值均<0.01)。BI-D1870处理组Molt-4细胞的凋亡率及G2/M期细胞所占百分比明显高于对照组(未进行药物干预)(P<0.01,P<0.05);BI-D1870联合依托泊苷处理组Molt-4细胞的凋亡率明显高于BI-D1870和依托泊苷单药处理组(P值均<0.01)。结论:多种AL细胞中RSK1和RSK2 m RNA的表达水平上调;RSK抑制剂BI-D1870联合依托泊苷可协同抑制AL细胞的增殖,诱导细胞凋亡及阻滞细胞周期。

Objective： To investigate the effects of p90 ribosomal S6 kinase （RSK） inhibitor BI-D1870 on proliferation, apoptosis and cell cycle distribution of acute leukemia （AL） cells.lvlethods： The expression levels of RSK1 and RSK2 mRNAs in six AL cell lines （Molt-4, Jurkat, Nalm-6, THP1, U937 and NB-4） were detected by real-time fluorescent quantitative-PCR. The effect of RSK inhibitor BI-D1870 on proliferation of AL cells and the inhibitory effects of BI-D1870 and etoposide alone and the combination of BI-D1870 and etoposide on the proliferation of Molt-4 cells were examined by CCK-8 assay. The apoptosis and cell cycle distribution of Molt-4 cells after treatment with BI-D1870 and the effects of BI-D1870 and etoposide alone and the combination of BI-D1870 and etoposide on the apoptosis of Molt-4 cells were detected by flow cytometry.Results： The expression levels of RSK1 mRNA in Molt-4 and THPlcell lines and the expression levels of RSK2 mRNA in Molt-4, Jurkat, THP1, U937 and NB-4 cell lines were higher than those in peripheral blood mononuclear cells from healthy volunteers （all P 〈 0.05）. The proliferation of Molt-4, Jurkat, Nalm-6, THP1, U937 and NB-4 cell lines were inhibited by RSK inhibitor BI-D1870 in a time-dependent manner （P 〈 0.05）. The proliferation inhibition rate of Molt-4 cells treated with the combination of BI-D1870 and etoposide was higher than those of the cells treated with BI-D1870 and etoposide alone （both P 〈 0.01）. The apoptosis rate and the percentage of cells in G2/M stage of Molt-4 cells treated with BI-D1870 were higher than those of the cells without treatment （P 〈 0.01, P 〈 0.05）. The apoptosis rate of Molt-4 cells treated with the combination of BI-D1870 and etoposide was higher than those of the cells treated with BI-D1870 and etoposide alone （both P 〈 0.01）.Conclusion： The mRNA expression levels of RSK1 and RSK2 are up-regulated in selected AL cells. RSK inhibitor BI-D1870 in combination with etoposide can synergistically inhibit the proliferation and induce the apoptosis of AL cells with cell cycle arrest.