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目的检测核干细胞因子(Nucleostemin,NS)基因在急性白血病细胞的表达情况,探讨NS基因与急性白血病发病机制之间的关系。方法从NS基因的3个变异体中筛选出共同序列设计特异性引物,应用逆转录-聚合酶链式反应技术(RT-PCR)对白血病细胞系K562、HL60和急性粒细胞白血病(M1、M2a和M3),急性单核细胞白血病(M5a和M5b),急性淋巴细胞白血病(ALL)进行NS基因表达水平的检测,以健康人和良性贫血患者骨髓单个核细胞作为对照。PCR产物片段为418bp。结果K562、HL60明显高表达NS基因,它与内参相比灰度值分别为0·735±0·260、0·449±0·190;急性白血病病例有相似的结果。M1+M2a、M3、M5a、M5b、ALL的灰度值分别为0·687±0·210、0·408±0·160、0·866±0·270、0·448±0·190、0·403±0·190;对照组健康人和良性贫血不表达或极微弱表达NS基因;同一细胞类型白血病,细胞分化停滞在早期阶段的表达NS基因的水平明显高于细胞分化停滞在后期的白血病,如K562高于HL60;M1、M2a高于M3;M5a高于M5b(P<0·01)。结论NS基因在急性白血病细胞中呈高表达状态,这与白血病的发生、发展有一定关联;不同分化阶段的白血病细胞表达NS基因水平存在着差异。NS基因在急性白血病细胞的表达情况也显示了癌细胞和干细胞在某些方面的相似性,可能是一个潜在的基因治疗靶位。
Objective To detect the expression of nucleostemin (NS) gene in acute leukemia cells and to explore the relationship between the NS gene and the pathogenesis of acute leukemia. Methods Specific primers designed by common sequences were screened from three variants of NS gene. The leukemia cell lines K562, HL60 and acute myeloid leukemia (M1, M2a) were reverse transcriptase-polymerase chain reaction (RT-PCR) And M3), acute monocytic leukemia (M5a and M5b), acute lymphoblastic leukemia (ALL) NS gene expression levels in healthy people and patients with benign anemia bone marrow mononuclear cells as a control. The PCR product fragment was 418 bp. Results The expression of NS gene in K562 and HL60 cells was significantly higher than that in control samples. The gray values of K562 and HL60 were 0. 735 ± 0. 260 and 0. 449 ± 0. 190, respectively. The acute leukemia patients had similar results. The grayscale values of M1 + M2a, M3, M5a, M5b and ALL are respectively 0.687 ± 0.210, 0.408 ± 0.160, 0.86 ± 0.270 and 0.448 ± 0.190 · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · + , Such as K562 higher than HL60; M1, M2a higher than M3; M5a higher than M5b (P <0.01). Conclusion NS gene is highly expressed in acute leukemia cells, which is correlated with the occurrence and development of leukemia. There are differences in NS gene expression in leukemia cells at different differentiation stages. The expression of NS gene in acute leukemia cells also shows some similarities between cancer cells and stem cells, which may be a potential gene therapy target.