摘要
目的探讨含同源2结构域蛋白酪氨酸磷酸酶-2(SHP-2)激活突变对人脑胶质瘤细胞凋亡及增殖活性的影响,为抑制肿瘤恶性转变寻找新的靶点。方法设计载有pc DNA3.1SHP-2D61Gmutant激活突变基因的质粒转染U521胶质瘤细胞,将稳定表达SHP-2的细胞作为为突变组,设置不含激活突变基因的空载质粒转染的细胞为空载组和未转染细胞作为正常对照组。在相同条件下对3组细胞进行培养,测定各组细胞增殖活性,侵袭能力以及细胞凋亡情况,评价脑胶质瘤细胞凋亡及增殖活性。采用t检验和c^2检验,组间的多重比较采用SNK-q检验进行统计学分析。结果培养24 h后突变组转染细胞增殖速度及活力相较于对照组和空载组均显著升高(t=13.236,16.782,P均<0.01);突变细胞侵袭能力(431.25±4.98)相较于对照组(66.05±5.13)和空载组(67.11±5.25)均明显升高(t=22.343,26.112,P均<0.01),细胞凋亡率(8.23﹪)相较于对照组(33.76﹪)和空载组(26.77﹪)也均显著降低(t=32.267,22.982,P均<0.01)。结论 SHP-2激活突变可提高神经胶质瘤细胞增殖速度、侵袭能力和细胞活力,发生恶性生物学行为改变,可能成为抗肿瘤恶性转变的新靶点。
Objective To investigate the effect of SH2 domain-containing protein-tyrosine phosphatase-2 （SHP-2） activation mutation on apoptosis and proliferation of human glioma cells, and to find a new target for inhibiting malignant transformation. Methods The design of pcDNA3.1 loaded SHP-2D61Gmutant activating mutation plasmid was transfected into U521 glioma cells, and the cells stably expressing SHP-2 were used as a mutation group. The cells transfected with empty plasmid without activating mutation gene were regarded as an empty group, and non-transfected cells served as a normal control group. Three groups of cells were cultured under the same condition. And the cell proliferation, invasion and cell apoptosis was measured to evaluate the apoptosis and proliferation of glioma cells. The t test and Z2 test were used as statistical analysis, and multiple comparisons between groups were performed by SNK-q test. Results Compared with the normal control group and empty vector group, the proliferation rate and activity of the transfected cells were significantly increased after 24 h （t = 13.236,16.782, P 〈 0.01 ） ; the invasion ability of the mutant cells was significantly increased [（431.25±4.98）vs （66.05 ±5.13）, （t = 22.343, P 〈 0.01 ） ; （431.25 ± 4.98 ） vs （ 67.11± 5.25 ）, （t = 26.112, P 〈 0.01 ） ], and the apoptosis rate was significantly reduced （8.23 % vs 33.76 %, 8.23 % vs 26.77 %, t = 32.267, 22.982, P 〈 0.01 ）. Conclusion SHP-2 activating mutation could improve the proliferation rate, invasion ability and cell viability of glioma cells, and change the malignant biological behavior. The result suggested that this protein may be a new target for malignant transformation.
出处
《中华细胞与干细胞杂志（电子版）》
2017年第4期214-218,共5页
Chinese Journal of Cell and Stem Cell（Electronic Edition）
