γ射线诱发的白血病小鼠骨髓细胞中SHP-2 mRNA及蛋白表达的变化

Changes of SHP-2 mRNA and protein of bone marrow cells of mice with leukemia induced by γ-ray irradiation

目的探讨蛋白酪氨酸磷酸酶2(Src homology 2 dom ain-contain ing tyrosine phosphatase 2,SHP-2)mRNA表达、蛋白表达及其功能在γ射线体外诱发的白血病小鼠骨髓细胞中的变化情况,为进一步从细胞信号转导途径入手,研究辐射致癌的分子机制奠定一定的基础。方法实验分为白血病组、辐射未癌变组及对照组,应用荧光定量PCR法、W est-ern b lot法及生物化学方法分别检测各组骨髓有核细胞中SHP-2 mRNA、蛋白表达量及其酶活性的变化情况。结果白血病组、辐射未癌变组及对照组SHP-2 mRNA含量分别为(5.08±2.87)、(4.59±2.36)、(3.54±1.02);SHP-2的蛋白表达水平(以密度定量的光密度表示)分别为(0.956±0.125)、(0.892±0.236)、(0.712±0.368);SHP-2的酶催化活性(以吸光度表示)分别为(0.156±0.069)、(0.118±0.065)及(0.098±0.048)。各组间差异无显著性。结论在γ射线诱发的白血病小鼠骨髓细胞中,SHP-2 mRNA含量、蛋白表达量及其功能无明显变化。

Objective To investigate alteration of SHP-2 mRNA and protein of bone marrow cells of leukemia mice after γ-ray exposure. Methods A total of 318 BALB/c mice were exposed to ^60Co γ-ray once a week for 4 weeks, and the total dose of γ-ray received by mice was 7.00 Gy. By pathological examination, 39 mice developed thymic lymphoma, 14 acute lymphoblastic leukemia, 21 T-lymphoblast leukemia/lymphoma, 1 spiroma, 4 malignant yolk sac tumors. Exposed to γ-ray, the mice that developed leukemia or were free of canceration were used in our study （n = 10 in each group） , and another 10 mice free from irradiation served as control. SHP-2 mRNA and protein in femoral bone marrow cells were detected by real time fluorescence quantitative PCR （FQ-PCR） and Western blotting respectively. Results SHP-2 mRNA was （5.08 ± 2. 87） in leukemia mice, （4. 59 ± 2. 36 ） in mice free of canceration and （3.54 ± 1.02） in controls. SHP-2 protein was （0. 956±0. 125） in leukemia mice, （0. 892 ±0. 236） in mice free of canceration and （0. 712 ±0. 368） in controls. The enzyme catalytic activity of SHP-2 was （0. 156±0. 069）, （0. 118 ±0. 065）, （0. 098 ±0. 048）. No significant difference in mRNA, protein or enzyme catalytic activity of SHP-2 was found in leukemia mice, mice free from canceration and control mice. Conclusion SHP-2 mRNA and protein was significantly increased in bone marrow cells of leukemia mice induced by γ-ray irradiation.