论文部分内容阅读
目的探讨蛋白酶体抑制剂硼替佐米单药以及联合柔红霉素对成人急性白血病原代细胞生长、凋亡和Bcl-2 mRNA表达的影响。方法用Ficoll液分离各型成人急性白血病患者骨髓单个核细胞,分别加入不同浓度硼替佐米(5、10、20、50 nmol/L)、柔红霉素(50、100、200、500 nmol/L),以及低浓度硼替佐米(5、10 nmol/L)联合不同浓度柔红霉素(50、100、200、500 nmol/L)进行细胞培养;采用MTT方法检测培养48 h后细胞增殖活性,计算抑制率、半抑制浓度(IC50)和相互作用系数(CDI)。流式细胞术检测培养24 h后细胞凋亡率,RT-PCR检测培养48h后凋亡基因Bcl-2 mRNA表达。结果所测各型急性白血病细胞生长抑制率随硼替佐米或柔红霉素单药浓度增高而增高;柔红霉素联合小浓度硼替佐米(5、10 nmol/L)后,柔红霉素的IC50由(102±27)nmol/L分别减小为(73±26)、(55±22)nmol/L;柔红霉素200 nmol/L联合硼替佐米10 nmol/L时协同作用最为显著,CDI=0.17。柔红霉素100 nmol/L联合硼替佐米20 nmol/L组与未加药组或单药组比较细胞凋亡率增高、Bc-l2 mRNA表达下降(P<0.05)。结论硼替佐米联合柔红霉素对成人急性白血病原代细胞有协同促凋亡和抑制白血病细胞生长的作用,有一定的临床应用前景。
Objective To investigate the effect of proteasome inhibitor bortezomib combined with daunorubicin on the growth, apoptosis and Bcl-2 mRNA expression of adult acute leukemia cells. Methods Bone marrow mononuclear cells from patients with acute leukemia of various types were isolated by Ficoll liquid and treated with different concentrations of bortezomib (5, 10, 20 and 50 nmol / L), daunorubicin (50, 100, 200 and 500 nmol / L, and low concentrations of bortezomib (5,10 nmol / L) combined with different concentrations of daunorubicin (50,100,200,500 nmol / L) for cell culture; MTT method was used to detect the cell proliferation after 48 h Activity, calculated inhibition, half-inhibitory concentration (IC50) and interaction coefficient (CDI). Flow cytometry was used to detect the rate of apoptosis after 24 h culture, and the expression of Bcl-2 mRNA was detected by RT-PCR 48 h after culture. Results The growth inhibition rates of all kinds of acute leukemia cells were increased with the increase of single dose of bortezomib or daunorubicin. After daunorubicin combined with low concentration of bortezomib (5 and 10 nmol / L) (IC50) decreased from (102 ± 27) nmol / L to (73 ± 26) and (55 ± 22) nmol / L respectively; synergistic effect of 200 nmol / L daunorubicin and 10 nmol / L bortezomib The most significant, CDI = 0.17. The apoptosis rates of daunorubicin 100 nmol / L and bortezomib 20 nmol / L group were significantly higher than those of the untreated or monotherapy group (P <0.05). Conclusion Bortezomib combined with daunorubicin has the synergistic effect of promoting apoptosis and inhibiting the growth of leukemia cells in adult acute leukemia primary cells and has certain clinical application prospects.