【摘要】 目的研究Shp-2酪氨酸蛋白磷酸酶对白血病细胞恶性增殖的影响。方法应用W estern b lot和RT-PCR技术分析白血病细胞和正常造血细胞样本Shp-2蛋白和mRNA表达水平;采用反义寡核苷酸技术和流式细胞术分析下调Shp-2表达对白血病细胞增殖与凋亡的影响。结果与正常造血细胞相比,磷酸化Shp-2蛋白在白血病细胞中呈过度表达状态,Shp-2与-βactin比值在白血病组(n=36)、正常骨髓造血细胞组(n=8)及正常外周血细胞组(n=12)分别为1.23±0.74、0.163±0.088和0.034±0.045。同样,Shp-2 mRNA在白血病细胞样本中呈高表达状态,其与-βactin比值在白血病组(n=26)和正常造血细胞组(n=25)分别为1.242±1.057和0.046±0.076。用Shp-2特异性反义寡核苷酸阻断Shp-2表达后,白血病细胞出现明显凋亡和生长抑制。结论Shp-2在人白血病细胞中呈过度表达状态,其表达水平可能与白血病细胞增殖程度密切相关。更多还原

【Abstract】 Objective To investigate the effects of Shp-2 tyrosine phosphatase activity on unlimited proliferation of leukemia cells.Methods Shp-2 protein and mRNA were evaluated with Western blot and RT-PCR,respectively.Shp-2 specific antisense was used to block Shp-2 expression in leukemia cells and the effects of down-regulation of Shp-2 on the growth and apoptosis of leukemia cells were determined by clone forming assay and flow cytometry(FCM).Results Shp-2 protein was overexpressed in most primary leukemia cell samples from various subtypes of leukemias,as compared to normal hematopoietic cells.The mean ratios of Shp-2/β-actin in leukemia samples(n=36),normal BM(n=8) and PBMC samples(n=12) were 1.23±0.74,0.163±0.088 and 0.034±0.045,respectively.Likely,high levels of Shp-2 mRNA were also observed in most leukemia cases,and the mean ratios of Shp-2/β-actin in leukemia cases(n=26) and PBMCs from healthy donors(n=25) were 1.242±1.057 and 0.046±0.076,respectively.Down-regulation of Shp2 expression caused remarkbaly apoptosis and inhibition of leukemic cell growth.Conclusions Shp-2 tyrosine phosphatase is overexpressed in most adult leukemia cases,and its activity positively correlates with proliferative potential of leukemia cells.更多还原