糖原合酶激酶-3β调节 U87-MG 人神经胶质瘤细胞的星形胶质细胞分化。
Glycogen synthase kinase-3beta regulates astrocytic differentiation of U87-MG human glioblastoma cells.
机构信息
Department of Infectious Diseases, Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
出版信息
Acta Pharmacol Sin. 2010 Mar;31(3):355-60. doi: 10.1038/aps.2010.10. Epub 2010 Feb 15.
AIM
To evaluate the role of glycogen synthase kinase-3beta (GSK-3beta) in the induced differentiation of human glioblastoma cells.
METHODS
Cell proliferation was determined by bromodeoxyuridine (BrdU) incorporation assay. The protein level of p-GSK-3beta, GSK-3beta, glial fibrillary acidic protein (GFAP) and proliferating cell nuclear antigen (PCNA) were determined using Western blots. The overexpression of mutant GSK-3beta was analyzed by immunocytochemistry.
RESULTS
The biotoxin cholera toxin is capable of inducing differentiation of U87-MG human glioblastoma cells, which is characterized by morphological changes to astrocytic phenotype, increase in differentiation marker protein GFAP and decrease in proliferation. GSK-3beta activation is induced during this differentiation. Small interfering RNA against GSK-3beta suppresses the induced-differentiation in U87-MG cells. Conversely, overexpression of a constitutively active form of human GSK-3beta (pcDNA3-GSK-3beta-S9A) mutant leads to differentiation of U87-MG cells.
CONCLUSION
Our findings suggest that GSK-3beta plays an important role in astrocytic differentiation of human glioblastoma cells and may be a novel therapeutic target in the malignant tumor.
目的
评估糖原合酶激酶-3β（GSK-3β）在人胶质母细胞瘤细胞诱导分化中的作用。
方法
通过溴脱氧尿苷（BrdU）掺入试验测定细胞增殖。采用 Western blot 法测定磷酸化 GSK-3β、GSK-3β、胶质纤维酸性蛋白（GFAP）和增殖细胞核抗原（PCNA）的蛋白水平。免疫细胞化学分析突变型 GSK-3β的过表达。
结果
生物毒素霍乱毒素能够诱导 U87-MG 人胶质母细胞瘤细胞分化，其特征为向星形胶质细胞表型的形态变化、分化标志物蛋白 GFAP 增加和增殖减少。在此分化过程中诱导 GSK-3β 激活。针对 GSK-3β 的小干扰 RNA 抑制 U87-MG 细胞的诱导分化。相反，过表达人 GSK-3β（pcDNA3-GSK-3β-S9A）突变的组成性激活形式导致 U87-MG 细胞分化。
结论
我们的发现表明 GSK-3β 在人胶质母细胞瘤细胞的星形胶质细胞分化中起重要作用，可能是恶性肿瘤的新治疗靶点。
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