CALM/AF10反义核酸对原代CALM/AF10融合转录白血病细胞的体外作用分析

Effects of CALM /AF10 antisenses on primary leukemic cells with CALM /AF10 fusion transcripts in vitro

目的 :确定 CALM和 AF1 0融合转录在原发性 t( 1 0 ;1 1 )转换的白血病形成和治疗中的作用。方法 :检测 5例原发性 t( 1 0 ;1 1 )转换的白血病患者 CAL M- AF1 0融合转录 ,检测 CALM/AF1 0硫代反义核酸对其白血病细胞体外化疗敏感性和凋亡的作用。结果 :RT-PCR在 5例原发性 t( 1 0 ;1 1 )转换的白血病患者中检测到 5种分子量大小的 AF1 0 - CALM融合转录和 4种分子量大小的 CAL M- AF1 0融合转录。原代 t( 1 0 ;1 1 )转换的白血病细胞对体外化疗敏感性较非 t( 1 0 ;1 1 )转换的显著低 ,而 AS- PS- ODNs增加了其体外化疗敏感性和凋亡率。5μM AS- PS- ODNs处理时有 4例阳性 ,1 0μM和 2 0μM AS- PS- ODNs处理时 5例全阳性 ,而单纯化疗药组和化疗药 + 1 0μM正义硫代核酸 ( S- PS- ODNs)组分别只有 3例。化疗药+ 1 0μM AS- PS- ODNs处理 48h、72 h、96h后 ,细胞凋亡指数分别显著高于单纯化疗药组和化疗药 + 1 0μM S- PS- ODNs组。而单纯化疗药组和化疗药 + 1 0μM S- PS- ODNs组间凋亡指数在对应时间无差别。结论 :CALM- AF1 0与 t( 1 0 ;1 1 )转换白血病的发生密切相关 ,且 t( 1 0 ;1 1 )转换的白血病患者预后较差。

Objectives: To investigate the in volvement of CALM and AF10 fusion transcripts in primary leukaemias with t(10;11). Methods: We detected the AF10 and CALM fusion in five t(10;11) leukemia samples by reverse transcriptase-poly merase chain reaction (RT-PCR), and effects of CALM/AF10 antisense phosphorothi oate oligodeoxynucleotides (AS PS-ODNs) on its chemotherapy sensitivity in vitr o and apoptosis. Results: Five different-sized AF10-CALM products and four dif ferent-sized CALM-AF10 products were detected by RT-PCR. The chemotherapy sen sitivity of leukemic cells with t(10;11) to drugs in vitro is lower than that of leukemic cells without t(10;11). AS PS-ODNs increased its chemotherapy sensiti vity and apoptotic rate. There were 4 cases positive at 5 μM concentration, all cases positive at 10 μM and 20μM concentration, vs only chemotherap eutic drugs (3 cases positive), and chemotherapeutic drugs + S-PS-ODNs (10 μM ) (3 cases positive). After cells were treated with 10 μmol/ L AS-PS-ODNs + c hemotherapeutic drugs for 48 h, 72 h, 96 h, the apoptotic indexis were 14.22±2. 8 6, 29.39 ±3.57, and 41. 26±4.52, respectively. These were significantly higher than that of only chemotherapeutic drugs-treated cells, and that of chemothera peutic drugs + S-PS-ODNs-treated cells at corresponding time. There was no d ifference between only drugs group and S-PS-ODNs group at corresponding time . Conclusion: The CALM and AF10 fusion transcript is involv ed in the pathogenesis of haematological malignancies with t(10;11) , and is associated with a poor prognosis.