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目的通过研究HL-60细胞在多种化疗药物作用后mRNA和蛋白的改变,初步探讨Survivin基因和耐药性之间的关系。方法用合适浓度的柔红霉素(DNR)、米托蒽醌(MIT)和三氧化二砷(As2O3)作用于HL-60细胞,随后用RT-PCR检测给与药物作用的第1天和第3天HL-60细胞中Survivin mRNA的表达情况,用Western blot检测Survivin蛋白的表达情况。结果在给予化疗药的第1天,3组HL-60细胞中Survivin mRNA表达水平全部降低:其中DNR下降了10％,MIT下降40％(P< 0．01),As2O3下降了25％(P<0．01)。在第3天,给予DNR和MIT组的HL-60细胞中Survivin mRNA表达较第1天有明显的升高, 分别上升20％(P<0．05)和65％(P<0．01);但给予As2O3组的Survivin mRNA表达仍持续降低,仅为第1天的68％(P<0．01)。 Western blot检测发现给予DNR和MIT作用3天后的HL-60细胞中Survivin蛋白含量分别升高了14％和11％,而给予As2O3组则下降了82％。结论白血病细胞给予化疗药后Survivin表达降低然后升高的现象可能与化疗中耐药性的发生有关;而三氧化二砷因有着不同的作用机制,可能会在逆转耐药性上发挥重要的作用。
OBJECTIVE: To investigate the relationship between Survivin gene and drug resistance by studying the changes of mRNA and protein in HL-60 cells treated with various chemotherapeutics. Methods HL-60 cells were treated with appropriate concentrations of daunorubicin (DNR), mitoxantrone (MIT) and arsenic trioxide (As2O3), followed by RT-PCR on days 1 and 3 The expression of Survivin mRNA in HL-60 cells was detected by Western blot. Results On the first day of chemotherapy, the expression of Survivin mRNA in HL-60 cells was significantly decreased in all the three groups (DNR decreased 10%, MIT decreased 40% (P <0.01) and As2O3 decreased 25% (P <0.01). On day 3, the expression of Survivin mRNA in HL-60 cells treated with DNR and MIT increased significantly by 20% (P <0.05) and 65% (P <0.01), respectively, ; However, the expression of Survivin mRNA in As2O3 group continued to decline, only 68% of the first day (P <0.01). Western blot showed that the protein levels of Survivin in HL-60 cells increased by 14% and 11%, respectively, after treated with DNR and MIT for 3 days, but decreased by 82% in As2O3 group. Conclusions The decrease and then increase of Survivin expression after leukemia cells receiving chemotherapeutic drugs may be related to the development of drug resistance in chemotherapy. However, arsenic trioxide may play an important role in reversing drug resistance because of its different mechanism.