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目的研究Src癌基因同源的酪氨酸蛋白磷酸酶2(Shp2)在慢性粒细胞白血病(CML)细胞中的表达,及其在p210bcr/abl诱导的白血病细胞恶性增殖和凋亡抵抗中的作用。方法收集25例p210bcr/abl阳性CML患者白血病细胞样本,8例非肿瘤病人骨髓和10例正常人外周血细胞样本作阴性对照,K562和KU812白血病细胞系作为p210bcr/abl阳性对照,KG1白血病细胞作为Shp2阳性对照。用Western印迹技术定量分析与比较Shp2在白血病细胞和正常骨髓造血细胞中的表达情况,通过特异性抑制剂分别下调Shp2和白血病融合基因p210bcr/abl后,观察Shp2表达对p210bcr/abl阳性白血病细胞增殖与凋亡的作用。细胞增殖与凋亡检测应用流式细胞仪。结果(1)磷酸化Shp2蛋白在92%患者CML白血病细胞样本中呈高表达状态,而在8例非肿瘤患者骨髓和10例正常人外周血细胞中低表达或不表达。磷酸化Shp2/β肌动蛋白比值分别为0.91±0.62、0.16±0.09和0.03±0.05(P均<0.01)。(2)下调Shp2蛋白表达后,白血病细胞凋亡率从4.89%上升到38.69%(P<0.01),而S期细胞从33.6%下降到10.8%(P<0.01)。(3)特异性抑制p210bcr/abl融合基因表达蛋白后,白血病细胞中磷酸化Shp2蛋白明显下降,同时出现明显细胞凋亡与生长抑制现象。结论(1)磷酸化Shp2蛋白在慢性粒细胞白血病细胞患者中呈过度表达状态,并且与白血病细胞恶性增殖与凋亡抵抗密切相关。(2)bcr/abl融合基因阳性患者的白血病细胞中Shp2的过度表达可能由p210bcr/abl蛋白激活引起。
Objective To investigate the expression of Src oncogene tyrosine phosphatase 2 (Shp2) in chronic myeloid leukemia (CML) cells and its role in p210bcr / abl-induced leukemic cell proliferation and apoptosis . Methods Twenty-five leukemia cell samples of p210bcr / abl positive CML patients were collected. Eight non-tumor bone marrow samples and 10 normal peripheral blood samples were collected as negative control. K562 and KU812 leukemia cell lines were used as p210bcr / abl positive control. KG1 leukemia cells were used as Shp2 Positive control. The expression of Shp2 in leukemia cells and normal bone marrow hematopoietic cells was quantitatively analyzed and compared by Western blotting. Shp2 and leukemia fusion gene p210bcr / abl were respectively down-regulated by specific inhibitors, and the effect of Shp2 expression on the proliferation of p210bcr / abl positive leukemia cells was observed And the role of apoptosis. Cell proliferation and apoptosis detection using flow cytometry. Results (1) Phospho-Shp2 protein was overexpressed in 92% of patients with CML leukemia cells, whereas 8 cases of non-tumor bone marrow cells and 10 normal human peripheral blood cells showed low or no expression. The phosphorylated Shp2 / β actin ratios were 0.91 ± 0.62, 0.16 ± 0.09 and 0.03 ± 0.05, respectively (all P <0.01). (2) The apoptosis rate of leukemia cells increased from 4.89% to 38.69% (P <0.01), while the S phase cells decreased from 33.6% to 10.8% (P <0.01) after down-regulation of Shp2 protein expression. (3) Phosphorylation of Shp2 protein in leukemia cells was significantly decreased after specific inhibition of p210bcr / abl fusion gene expression, accompanied with obvious apoptosis and growth inhibition. Conclusion (1) Phospho-Shp2 protein is overexpressed in patients with chronic myeloid leukemia and is closely related to the proliferation and apoptosis of leukemic cells. (2) The overexpression of Shp2 in leukemia cells with bcr / abl fusion gene positive may be caused by the activation of p210bcr / abl protein.