论文部分内容阅读
本研究探讨重组人粒细胞刺激因子G-CSF联合小剂量阿糖胞苷(cytarabine,Ara-C)和高三尖杉酯碱(homoharringtonine,HHT)GHA预激化疗方案治疗复发难治、老年、低增生急性单核细胞白血病的临床疗效及其不良反应,同时以U937细胞株作为体外模型,初步探讨GHA预激方案的作用机制。对37例AML-M5患者采用GHA预激化疗方案,具体为G-CSF200μg/(m2.d),皮下注射,第0-14天;高三尖杉酯碱1mg/(m2.d),静脉点滴,第1-14天;阿糖胞苷10mg/m2,皮下注射,每12小时1次,第1-14天。观察临床疗效、不良反应、治疗相关死亡率等。选择U937细胞株为体外模型,观察G-CSF作用对细胞周期的影响,不同浓度的含G-CSF的预激方案(GHA)及不含G-CSF的常规诱导缓解方案(HA)对细胞形态、抑制率、凋亡的作用;用免疫组织化学方法检测药物作用前后U937细胞MLAA34蛋白表达情况。结果表明,37例AML-M5患者中,总有效率为62.2%,其中CR患者占45.95%(17/37),PR患者16.22%(6/37);中性粒细胞缺乏18.92%(7/37),中位时间4天;重症肺部感染2例;无明显出血、消化道反应,无神经肾毒性发生。U937细胞经G-CSF作用24小时后,S期细胞比例显著增高;在GHA和HA方案作用下细胞生长受抑制,形态上可观察到凋亡表现;流式细胞仪也可测出有早期凋亡,MLAA34蛋白表达与未经药物作用的细胞相比,表达降低。GHA和HA组相比较,细胞抑制率、早期凋亡率及MLAA34蛋白表达下调情况均有显著差异,有统计学意义。结论:①含G-CSF联合小剂量Ara-C和HHT的GHA预激化疗方案是一种高效低毒的化疗方案,用于治疗难治复发、老年及低增生AML-M5效果肯定,血液系统和非血液系统不良反应发生率低。②GHA预激方案可能机制为G-CSF刺激细胞S期比例增高,进入细胞周期,从而增强细胞周期特异性化疗药物毒性。GHA和HA方案均可抑制细胞增殖,诱导凋亡,但以GHA方案作用更为显著。③MLAA34是1个与AML-M5发生发展相关的抗凋亡分子。G-CSF联合HA的预激化疗方案可下调MLAA34蛋白的表达。
This study was to investigate the effect of recombinant human granulocyte-stimulating factor G-CSF combined with low-dose cytarabine (Ara-C) and homoharringtonine (HHT) GHA pretreatment on refractory relapsed, elderly, Proliferation of acute monocytic leukemia clinical efficacy and adverse reactions, at the same time to U937 cell line as an in vitro model to initially explore the mechanism of GHA pre-excitation program. In 37 AML-M5 patients, GHA chemoradiation regimen was used, specifically G-CSF 200 μg / (m2.d), injected subcutaneously on days 0-14, homoharringtonine 1 mg / (m2.d), intravenous drip , Day 1-14; Cytarabine 10 mg / m2, subcutaneously, every 12 hours, day 1-14. To observe the clinical efficacy, adverse reactions, treatment-related mortality. The U937 cell line was selected as an in vitro model to observe the effect of G-CSF on the cell cycle. Different concentrations of GHA with G-CSF and conventional induction remission without HA (G-CSF) , Inhibition rate and apoptosis. The expression of MLAA34 protein in U937 cells before and after drug treatment was detected by immunohistochemistry. The results showed that the total effective rate was 62.2% in 37 AML-M5 patients, of which 45.95% (17/37) were CR patients, 16.22% (6/37) in PR patients and 18.92% (7 / 37), the median time of 4 days; 2 cases of severe pulmonary infection; no obvious bleeding, gastrointestinal reactions, no neurotoxicity nephrotoxicity. After treated with G-CSF for 24 hours, the proportion of cells in U937 cells was significantly increased. The cell growth was inhibited under the GHA and HA protocols, and morphological changes of apoptosis were observed. Flow cytometry also showed that there was early apoptosis The expression of MLAA34 protein was lower than that of the non-drug-treated cells. GHA and HA group compared to the cell inhibition rate, early apoptosis rate and MLAA34 protein expression were significantly different, with statistical significance. Conclusion: GHA combined with low dose of Ara-C and HHT GHA pre-shock chemotherapy program is a highly efficient and low toxicity chemotherapy for the treatment of refractory relapse, the elderly and low proliferative AML-M5 affirmation, the blood system Adverse reactions to non-hematologic malignancies were low. GHA pre-excitation program may mechanism for G-CSF stimulated cells increased the proportion of S phase into the cell cycle, thereby enhancing cell cycle-specific chemotherapy toxicity. Both GHA and HA regimens inhibited cell proliferation and induced apoptosis, but the effect was more pronounced with GHA. ③ MLAA34 is an anti-apoptotic molecule associated with the development of AML-M5. The pre-treatment regimen of G-CSF combined with HA downregulates MLAA34 protein expression.