PAX5/BSAP在儿童急性白血病细胞和血液肿瘤细胞株中的表达研究  

Expression of the transcription factor PAX5/BSAP in childhood acute leukemic cells and haematolological tumor cell lines

作　　者：张蓓[1] 袁向亮[1] 帖利军[2] 叶启东[2] 汤静燕[2] 沈立松[1] 

机构地区：[1]上海第二医科大学附属新华医院,上海儿童医学中心实验诊断中心,200127 [2]上海第二医科大学附属新华医院,上海儿童医学中心血液肿瘤科

出　　处：《中华检验医学杂志》2005年第11期1131-1135,共5页Chinese Journal of Laboratory Medicine

摘　　要：目的了解儿童急性白血病细胞和血液肿瘤细胞株中转录因子PAX5/BSAP的表达特性。方法采用实时定量RTPCR方法,检测了6个血液肿瘤细胞株以及6例正常儿童、58例初发和4例复发急性白血病儿童骨髓细胞中PAX5和CD19mRNA的表达水平。采用WesternBlotting分析了6个血液肿瘤细胞株和4个临床样本中BSAP表达水平。采用RNAi(RNA干扰)技术特异性阻断NAMALWA细胞株(B细胞系)中PAX5的表达。结果在NAMALWA细胞株中,PAX5和CD19mRNA相对表达量分别为2.35%和2.52%;而在T和髓系细胞株中几乎不表达。同时运用WesternBlotting在B系细胞中检测到一52KD的BSAP条带,但在T和髓系细胞中为阴性。在临床样本中BALL(急性B淋巴细胞白血病)组比TALL(急性T淋巴细胞白血病)组和AML(急性髓细胞性白血病)组的PAX5mRNA表达高(P=0.029和P=0.013)。在BALL患儿中,PAX5mRNA表达的个体差异很大。RNAi技术抑制NAMALWA细胞株中PAX5基因表达达35%,但不影响细胞增殖效率(P<0.05)。结论部分BALL中PAX5mRNA表达明显升高,可作为进一步研究BALL发病机理的另一个切入点。RNAi技术可进一步用于B系血液恶性肿瘤细胞的观察研究。Objective To investigate transcription factor PAXS/BSAP expression characteristics in childhood acute leukemic cells and haematolological tumor cell lines. Methods A real-time RT-PCR assay for the relative quantitation of PAX5 and CD19 mRNA expression was developed and applied on 6 haematological tumor cell lines and bone marrow cells of 6 normal children, 58 previously untreated and 4 relapse acute leukemic children. BSAP expression by Western Blotting analysis was performed in haematological tumor cells, including 6 haematological tumor cell lines and 4 clinical samples. RNAi （ RNA interference） was used to specifically block PAX5 expression in NAMALWA （B-cell lines）. Results PAX5 and CD19 mRNA expression level was 2. 35%, 2.52%, respectively, in NAMALWA, but almost not detectable in other T- and myeloid cell lines. The results of Western Blotting analysis showed a 52-KD BSAP band in B lineage cells, but not in T- and myeloid lineage cells. Among clinical samples, expression of PAX5 mRNA in B-ALL （acute B-lineage lymphoblastic leukemia） was significantly higher than that in T-ALL （acute T-lineage lymphoblastic leukemia） and AML （acute myeloid leukemia） （P = 0. 029 and P = 0. 013, respectively）. Individual difference of PAX5 mRNA expression levels in children with B-ALL was great. PAX5 suppression in NAMALWA by RNAi was 35%, but had no effect on cell proliferation （ P 〈 0. 05）. Conclusions The strong PAX5 mRNA expression in some B-ALL can be considered to be particularly interesting for further analysis. RNAi is also suitable for further investigation in B-lineage haematological tumor cells.

关 键 词：白血病 混合细胞 基因表达调控 白血病 

分 类 号：R733[医药卫生—肿瘤]