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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 the expression of PAX5 / BSAP in children with acute leukemia and hematologic malignancies. Methods Real-time quantitative RTPCR was used to detect the expression of PAX5 and CD19mRNA in 6 hematological tumor cell lines and 6 normal children, 58 primary and 4 recurrent acute leukemia children’s bone marrow cells. Western Blotting was used to analyze BSAP expression in six hematological tumor cell lines and four clinical samples. PAX5 expression in NAMALWA cell line (B cell line) was specifically blocked using RNAi (RNA interference) technique. Results In NAMALWA cell lines, the relative expression levels of PAX5 and CD19 mRNA were 2.35% and 2.52%, respectively. However, they were almost not expressed in T and myeloid cell lines. A 52KD BSAP band was detected in B line cells by Western Blotting, but negative in T and myeloid lineages. PAX5 mRNA expression was significantly higher in the BALL (acute B-lymphoblastic leukemia) group than in the TALL (acute lymphoblastic leukemia) and AML (acute myeloid leukemia) groups in clinical samples (P = 0.029 and P = 0.013). PAX5 mRNA expression varies widely among children with BALL. RNAi inhibited PAX5 gene expression in NAMALWA cells by 35%, but did not affect cell proliferation efficiency (P <0.05). Conclusions PAX5 mRNA expression in part of BALL is significantly increased, which may serve as another entry point for further study on the pathogenesis of BALL. RNAi technology can be further used for the observation of B hematological malignancies.