三氧化二砷和抗坏血酸对骨髓增生异常综合征患者 BCL2 家族基因的影响：一项体外研究。
Arsenic trioxide and ascorbic acid interfere with the BCL2 family genes in patients with myelodysplastic syndromes: an ex-vivo study.
机构信息
Department of Oncology, Transplant, New Advances in Medicine, Section of Hematology, University of Pisa, Pisa, Italy.
出版信息
J Hematol Oncol. 2012 Sep 10;5:53. doi: 10.1186/1756-8722-5-53.
BACKGROUND
Arsenic Trioxide (ATO) is effective in about 20% of patients with myelodysplasia (MDS); its mechanisms of action have already been evaluated in vitro, but the in vivo activity is still not fully understood. Since ATO induces apoptosis in in vitro models, we compared the expression of 93 apoptotic genes in patients' bone marrow before and after ATO treatment. For this analysis, we selected 12 patients affected by MDS who received ATO in combination with Ascorbic Acid in the context of the Italian clinical trial NCT00803530, EudracT Number 2005-001321-28.
METHODS
Real-time PCR quantitative assays for genes involved in apoptosis were performed using TaqMan® Assays in 384-Well Microfluidic Cards "TaqMan® Human Apoptosis Array".Quantitative RT-PCR for expression of EVI1 and WT1 genes was also performed. Gene expression values (Ct) were normalized to the median expression of 3 housekeeping genes present in the card (18S, ACTB and GAPDH).
RESULTS
ATO treatment induced up-regulation of some pro-apoptotic genes, such as HRK, BAK1, CASPASE-5, BAD, TNFRSF1A, and BCL2L14 and down-regulation of ICEBERG. In the majority of cases with stable disease, apoptotic gene expression profile did not change, whereas in cases with advanced MDS more frequently pro-apoptotic genes were up-regulated. Two patients achieved a major response: in the patient with refractory anemia the treatment down-regulated 69% of the pro-apoptotic genes, whereas 91% of the pro-apoptotic genes were up-regulated in the patient affected by refractory anemia with excess of blasts-1. Responsive patients showed a higher induction of BAD than those with stable disease. Finally, WT1 gene expression was down-regulated by the treatment in responsive cases.
CONCLUSIONS
These results represent the basis for a possible association of ATO with other biological compounds able to modify the apoptotic pathways, such as inhibitors of the BCL2 family.
背景
三氧化二砷（ATO）在约 20%的骨髓增生异常综合征（MDS）患者中有效；其作用机制已在体外进行了评估，但体内活性仍不完全清楚。由于 ATO 在体外模型中诱导细胞凋亡，我们比较了 12 名接受 ATO 联合抗坏血酸治疗的 MDS 患者骨髓中 93 个凋亡基因的表达。为此分析，我们选择了意大利临床试验 NCT00803530 中接受 ATO 联合抗坏血酸治疗的 12 名 MDS 患者，EudracT 编号为 2005-001321-28。
方法
使用 TaqMan® Assays 在 384 孔微流控卡“TaqMan® Human Apoptosis Array”上进行涉及凋亡的基因实时定量 PCR 定量检测。还进行了 EVI1 和 WT1 基因表达的定量 RT-PCR。基因表达值（Ct）通过中位数归一化校正存在于卡片中的 3 个管家基因（18S、ACTB 和 GAPDH）进行归一化。
结果
ATO 治疗诱导了一些促凋亡基因的上调，如 HRK、BAK1、CASPASE-5、BAD、TNFRSF1A 和 BCL2L14，以及 ICEBERG 的下调。在大多数疾病稳定的病例中，凋亡基因表达谱没有改变，而在 MDS 进展的病例中，促凋亡基因更常上调。两名患者取得了主要缓解：在难治性贫血患者中，治疗下调了 69%的促凋亡基因，而难治性贫血伴过多原始细胞-1 患者的 91%促凋亡基因上调。有反应的患者 BAD 的诱导明显高于疾病稳定的患者。最后，WT1 基因的表达在反应病例中受到治疗的下调。
结论
这些结果为 ATO 与其他能够改变凋亡途径的生物化合物的可能联合治疗提供了依据，如 BCL2 家族抑制剂。
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