Missense mutations in PML-RARA are critical for the lack of responsiveness to arsenic trioxide treatment
Missense mutations in PML-RARA are critical for the lack of responsiveness to arsenic trioxide treatment
复制标题
DOI：
10.1182/blood-2011-01-329433
复制
发表时间：
2011-08-11
期刊：
影响因子：
20.3
通讯作者：
Naoe, Tomoki
中科院分区：
文献类型：
--
作者：
Goto, Emi;Tomita, Akihiro;Naoe, Tomoki
三氧化二砷（As 2 O3）是治疗难治性/复发性急性早幼粒细胞白血病（APL）的一种高效治疗方法，但近年来出现了对As 2 O3的耐药性。在本研究中，我们报告的结果，2/15例难治性/复发性APL患者与As 2 O3治疗临床As 2 O3耐药。这2例患者的白血病细胞在早幼粒细胞白血病基因-维甲酸受体-α基因（PML-RARA）转录物中存在错义突变，导致PML B2结构域中A216 V和L218 P的氨基酸取代。当野生型或突变型PML-RARA（PR-WT和PR-B/L-mut，分别）在HeLa细胞中过表达，免疫印迹显示SUMO化和/或寡聚化蛋白带PR-WT，但不是在PR-B/L-mut后As 2 O3处理。蛋白质定位分析表明，PR-WT在可溶性组分转移到不溶性组分后，用As 2 O3处理，但PR-B/L-mut稳定检测馏分中有和没有As 2 O3。免疫荧光显微镜分析显示，PR-WT定位为细胞质中的微粒图案，没有As 2 O3和作为一个大颗粒图案与As 2 O3。PR-B/L-mut在有和没有As_2O_3的细胞质中广泛观察到。在患者的原代细胞中观察到几乎相同的定位模式。因此，B2结构域突变可能在对As 2 O3的异常分子反应中起重要作用，并且可能是APL中As 2 O3抗性的关键。（血。2011;118（6）：1600-1609）
Arsenic trioxide (As2O3) is a highly effective treatment for patients with refractory/relapsed acute promyelocytic leukemia (APL), but resistance to As2O3 has recently been seen. In the present study, we report the findings that 2 of 15 patients with refractory/relapsed APL treated with As2O3 were clinically As2O3 resistant. Leukemia cells from these 2 patients harbored missense mutations in promyelocytic leukemia gene-retinoic acid receptor-alpha gene (PML-RARA) transcripts, resulting in amino acid substitutions of A216V and L218P in the PML B2 domain. When wild-type or mutated PML-RARA (PR-WT and PR-B/L-mut, respectively) were overexpressed in HeLa cells, immunoblotting showed SUMOylated and/or oligomerized protein bands in PR-WT but not in PR-B/L-mut after As2O3 treatment. Protein-localization analysis indicated that PR-WT in the soluble fraction was transferred to the insoluble fraction after treatment with As2O3, but PR-B/L-mut was stably detected in fractions both with and without As2O3. Immunofluorescent microscopy analysis showed PR-WT localization as a microgranular pattern in the cytoplasm without As2O3 and as a macrogranular pattern with As2O3. PR-B/L-mut was diffusely observed in the cytoplasm with and without As2O3. Nearly identical localization patterns were observed in patients' primary cells. Therefore, B2 domain mutations may play an important role in aberrant molecular responses to As2O3 and may be critical for As2O3 resistance in APL. (Blood. 2011;118(6):1600-1609)