糖原合成激酶3β在骨肉瘤生长中的作用及其作用机制的研究

Osteosarcoma represents the malignant bone tumor that is most refractory to treatment and in which the identification of molecular targets is urgently required. Recently research indicates that glycogen synthase kinase-3beta (GSK3β), which is frequently associated with more and more research about osteosarcoma, may play a key role in the process of growth and proliferation of tumor cells. However, the mechanisms which cause these accommodations remain poorly understood. Our recent discovery that GSK3β represents overexpression and activity accommodation abnormal in osteosarcoma cells, prompted us to investigate whether GSK-3β inhibition alters growth processes of osteosarcoma. In this study, we will examine for GSK3β expression and its phosphorylation by Western immunoblotting and for GSK3βactivity by in vitro kinase assay.The effects of small-molecule GSK3βinhibitors on cell survival, proliferation, and apoptosis were examin in vitro and in vivo. In this research, we will clarificate that impact of GSK3β lose on stability adjustment of p27, cyclin-CDK-Rb pathway mediated the E2F transcription activit, hTERT gene expression and telomerase activity. The result will open up a potential target for clinical treatment of osteosarcoma.

骨肉瘤是恶性程度高且难治的骨肿瘤，寻求潜在的分子靶点是目前研究的焦点。糖原合成激酶3β(GSK3β)在骨肉瘤细胞中超表达且存在活性调节异常，提示GSK3β可能在骨肉瘤的生长中起正性调节作用。本研究为明确GSK3β在骨肉瘤中的正性调节作用，阐明GSK3β促进骨肉瘤生长的作用机制，拟使用Western blot法和自主开发的NIRKA法，检测骨肉瘤GSK3β的表达和活性；使用其抑制剂或siRNA减少内源性GSK3β表达或活性，在体内、外试验中检测骨肉瘤细胞的生存、增殖及肿瘤相关因子表达及磷酸化水平，初步阐明GSK3β对p27kip1蛋白稳定性、cyclin-CDK-Rb径路介导的E2F转录活性、hTERT基因表达和端粒酶活性的影响。GSK3β在骨肉瘤生长过程中发挥关键作用的证实，可以为骨肉瘤的治疗开辟新的靶点。

骨肉瘤是恶性程度高且难治的骨肿瘤，寻求潜在的分子靶点是目前研究的焦点。糖原合成激酶3β(GSK3β)在骨肉瘤细胞中超表达且存在活性调节异常，提示GSK3β可能在骨肉瘤的生长中起正性调节作用。本研究为明确GSK3β在骨肉瘤中的正性调节作用，阐明GSK3β促进骨肉瘤生长的作用机制，使用Western blot法和自主开发NIRKA法，检测骨肉瘤GSK3β的表达和活性；使用其抑制剂或siRNA减少内源性GSK3β表达或活性，在体内、外试验中检测骨肉瘤细胞的生存、增殖及肿瘤相关因子表达及磷酸化水平，初步阐明GSK3β对p27蛋白及其下游cyclinD1-CDK-Rb径路调控的分子机制。GSK3β在骨肉瘤生长过程中发挥关键作用的证实，可以为骨肉瘤的治疗开辟新的靶点。