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目的探讨急性白血病(AL)患者骨髓单个核细胞(BMMNC)视网膜母细胞瘤相关蛋白46(RbAp46)蛋白和基因表达状态,揭示RbAp46表达与AL类型及预后的可能联系。方法用Western印迹技术检测46例AL患者BMMNCRbAp46蛋白表达量;用RTPCR方法扩增22例AL患者BMMNCRbAp46mRNA的表达并进行半定量检测;用间接免疫荧光技术观察RbAp46蛋白在BMMNC的表达和分布。结果①AL患者BMMNC存在RbAp46蛋白和mRNA表达,表达强度与白血病类型无关。②肿瘤重度负荷组的AL患者BMMNC表达RbAp46蛋白的平均吸光度(A)值为93.4±37.2,明显低于肿瘤轻度负荷组的127.2±15.8(P<0.05)。③难治性白血病患者BMMNCRbAp46蛋白表达的平均A值为87.1±33.8,明显低于非难治性白血病组的126.6±21.2(P<0.05)。④肿瘤重度负荷组AL患者BMMNCRbAp46mRNA的相对表达量为0.19±0.08,明显低于肿瘤轻度负荷组的0.31±0.12(P<0.05)。⑤AL和对照组患者BMMNC均存在RbAp46蛋白的原位表达;RbAp46蛋白主要分布在细胞核中。结论AL患者BMMNC存在RbAp46蛋白和mRNA表达,该表达在肿瘤负荷重的AL和难治性白血病患者显著降低,提示RbAp46可能是白血病细胞生长的抑制基因。
Objective To investigate the protein and gene expression status of RbAp46 in bone marrow mononuclear cells (BMMNC) in patients with acute leukemia (AL) and to reveal the possible relationship between RbAp46 expression and AL type and prognosis. Methods The expression of BMMNCRbAp46 protein in 46 cases of AL patients was detected by Western blotting. The expression of BMMNCRbAp46 mRNA in 22 patients with AL was amplified by RTPCR and semi-quantitatively detected. The expression and distribution of RbAp46 protein in BMMNC were observed by indirect immunofluorescence staining. Results ① There was RbAp46 protein and mRNA expression in BMMNC of AL patients. The expression intensity of RbAp46 was not related to the type of leukemia. ② The mean absorbance (A) value of RbAp46 protein expression in BMMNC patients with severe tumor burden was 93.4 ± 37.2, which was significantly lower than 127.2 ± 15.8 (P <0.05) in mild tumor burden group. ③ The mean A value of BMMNCRbAp46 protein expression in patients with refractory leukemia was 87.1 ± 33.8, which was significantly lower than that in non-refractory leukemia patients (126.6 ± 21.2, P <0.05). ④ The relative expression of BMMNCRbAp46 mRNA in AL patients with severe tumor load was 0.19 ± 0.08, which was significantly lower than that of the mild tumor load group (0.31 ± 0.12, P <0.05). ⑤ RbAp46 protein was expressed in situ in BMMNC of AL and control group; RbAp46 protein mainly distributed in nucleus. Conclusion The expression of RbAp46 protein and mRNA in BMMNC of patients with AL is significantly lower than that in AL patients with refractory leukemia and tumor burden, suggesting that RbAp46 may be a suppressor of leukemia cell growth.