儿童急性淋巴细胞白血病及阿霉素所致Jurkat细胞凋亡时WAVE1表达研究

目的：初步探讨WASP家族Verprolin同源蛋白1（WAVE1）在急性淋巴细胞白血病（ALL）发病中可能的作用及意义。方法：运用逆转录-聚合酶链反应（RT-PCR）半定量法和蛋白质印迹（Western blotting）分别检测40例ALL初治患儿、15例化疗完全缓解半年、4例复发、10例非白血病患儿BMMCs中WAVE1 mRNA和蛋白的表达水平。以不同浓度阿霉素处理Jurkat细胞：①采用MTT法测细胞增殖；②采用流式细胞仪检测细胞凋亡率；③采用RT-PCR和Western blotting检测WAVE1表达。结果：ALL初治和复发患儿BMMCs均有WAVE1 mRNA和蛋白的表达，对照和缓解组BMMCs中mRNA和蛋白低表达或无表达,初治和复发组的WAVE1 mRNA和蛋白的表达表达水平显著高于化疗后缓解组和对照组（P<0.01）。阿霉素明显抑制Jurkat细胞增殖，抑制作用呈剂量时间依赖效应（P<0.05）；阿霉素作用24 h后，细胞凋亡随药物浓度增增高而增加，与对照组相比，差异有显著性(P<0.05)；细胞WAVE1 mRNA和蛋白表达水平随阿霉素处理浓度的增高而降低，与对照组比较，差异有显著性(P<0.05)。结论：WAVE1在ALL患儿BMMCs中高表达；WAVE1可能与儿童ALL病程相关，它可能成为动态检测儿童ALL病情的一个新指标。

xpression of WAVE1 in childhood acute lymphocytic leukemia and in the apoptosis of Jurkat cells induced by adriamycin

Objective To investigate whether WASP/Verprolin homologous protein 1(WAVE1)plays a role in the pathogenesis of childhood acute lymphoblastic leukemia(ALL).Methods WAVE1 mRNA and protein expression in bone marrow mononuclear cells(BMMCs)was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse,15 children with ALL in complete remission(CR)and 40 children with newly diagnosed ALL.Ten normal bone marrow samples were used as controls.Jurkat cells were treated with different concentrations of adriamycin(ADM).The cell proliferation was detected with MTT.The apoptosis rate was measured by flow cytometry.WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.Results WAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR.Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR(P<0.01).ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent(P<0.05).After ADM treatment for 24 hrs,the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls(P<0.05).Conclusions The WAVE1 expression increased in children with ALL.WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.