硼替佐米单独或联合三尖杉酯碱、三氧化二砷对耐药白血病细胞增殖、凋亡的影响

In vitro effect of bortezomib alone or in combination with harringtonine or arsenic trioxide on proliferation and apoptosis of mnitidrug resistant leukemia calls

目的探讨硼替佐米单独或联合三尖杉酯碱（HT）、三氧化二砷（As2O3）对HL-60／ADM多药耐药细胞株和难治、复发急性白血病原代细胞增殖、凋亡的影响。方法以HL-60／ADM白血病细胞株及难治、复发急性白血病患者原代细胞为研究对象，采用硼替佐米单独或联合HT、As2O3处理细胞，MTT法观察细胞增殖活力，Hoechst33342染色、流式细胞术检测细胞凋亡率及检测胞内阿霉素荧光阳性率，进一步分析硼替佐米逆转耐药能力。结果10～50nmol/L硼替佐米能够抑制HL-60／ADM细胞增殖并引起凋亡，其中40nmol/L处理48h达最大抑制效果；应用15μmol／L As2O3和752nmol/L HT分别与不同浓度硼替佐米联合处理HL-60／ADM细胞，其对HL-60／ADM细胞的增殖抑制率和凋亡率均高于硼替佐米单药处理组（P值均〈0．05）。10nmol／L硼替佐米能使HL-60／ADM细胞对阿霉素的蓄积作用大大提高。硼替佐米对难治、复发急性白血病患者原代细胞的增殖抑制作用与HL-60/ADM细胞一致。结论硼替佐米能够抑制HL60／ADM细胞及难治、复发急性白血病原代细胞增殖并诱导其凋亡，与HT或As2O3联合具有相加作用。

Objective To investigate the effect of bortezomib alone or combined with harringtonine （HT） or arsenic trioxide （As2O3 ） on the proliferation capacity and apoptosis of HL-60/ADM cell line and fresh cells from refractory/relapse acute leukemia patients. Methods HL-60/ADM cells or refractory/relapse leukemia cells were incubated with bortezomib at different doses alone and in combination with HT or As2 O3. The proliferation capacity was observed by MTT assay, cell apoptosis by fluorescence microscopy and flow cytometry. Intracellular concentration of dannotubicin （DNR） was determined by flow cytometry. Restilts In bortezomib-treated HL-60/ADM cells, the proliferation inhibition rate and apoptotic cells increased in a time- and dose-dependent manner. 40 nmol/L bortezomib could maximally inhibit the proliferation of HL- 60/ADM cells at 48 hours. 15 μmol/L As2O3 or 752 nmol/L HT combined with different doses of bortezomib could inhibit proliferation and induce apoptosis of HL-60/ADM cells. The As2O3 plus bortezomib or HT plus bortezomib showed a greater anticancer efficacy than either of the drugs alone （ P 〈 0.05, P 〈 0.01 ）. Bortezomib （10 nmol/L） could markedly enhance the intraeellular accumulation of DNR in HL-60/ADM cells （P 〈 0.05 ）. Conclusions Bortezomib can inhibit proliferation and induce apoptosis of HL-60/ADM cells and fresh refactory/relapse acute leukemia cells, especially combined with HT or As2O3.