Systematic Kinome shRNA Screening Identifies CDK11 (PITSLRE) Kinase Expression Is Critical for Osteosarcoma Cell Growth and Proliferation
Systematic Kinome shRNA Screening Identifies CDK11 (PITSLRE) Kinase Expression Is Critical for Osteosarcoma Cell Growth and Proliferation
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DOI：
10.1158/1078-0432.ccr-12-1157
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发表时间：
2012-09-01
影响因子：
11.5
通讯作者：
Hornicek, Francis J.
中科院分区：
文献类型：
--
作者：
Duan, Zhenfeng;Zhang, Jianming;Hornicek, Francis J.
目的：寻找新的靶向治疗方法是提高骨肉瘤患者生存率的关键。本研究的目的是确定骨肉瘤中基于激酶的潜在治疗靶点。实验设计：我们使用基于慢病毒的shRNA激酶文库来筛选在骨肉瘤细胞生存中发挥作用的激酶。采用细胞增殖实验检测细胞生长和存活情况。应用siRNA分析来证实所观察到的由激酶基因表达缺失引起的表型变化。CDK11(PITSLRE)是骨肉瘤细胞生存所必需的，Western印迹分析和免疫组织化学证实其表达。患者总生存期与CDK11的表达及预后相关。在骨肉瘤体内移植模型中进一步评价CDK11表达在维持骨肉瘤生长中的作用。结果：骨肉瘤细胞高水平表达CDK11。慢病毒shRNA或siRNA下调CDK11的表达可抑制骨肉瘤细胞的生长并诱导其凋亡。免疫组织化学分析显示CDK11高表达的骨肉瘤患者的生存期明显短于CDK11低表达的骨肉瘤患者。结论：CDK11信号通路在骨肉瘤细胞生长和存活中起重要作用，进一步阐明了CDK11表达的调控机制，并最终开发出一种CDK11抑制剂，可能对骨肉瘤有治疗作用。临床癌症资源；18(17)；4580-8。(C)2012年AACR。
Purpose: Identification of new targeted therapies is critical to improving the survival rate of patients with osteosarcoma. The goal of this study is to identify kinase based potential therapeutic target in osteosarcomas.Experimental Design: We used a lentiviral-based shRNA kinase library to screen for kinases which play a role in osteosarcoma cell survival. The cell proliferation assay was used to evaluate cell growth and survival. siRNA assays were applied to confirm the observed phenotypic changes resulting from the loss of kinase gene expression. CDK11 (PITSLRE) was identified as essential for the survival of osteosarcoma cells, and its expression was confirmed by Western blot analysis and immunohistochemistry. Overall patient survival was correlated with the CDK11 expression and its prognosis. The role of CDK11 expression in sustaining osteosarcoma growth was further evaluated in an osteosarcoma xenograft model in vivo.Results: Osteosarcoma cells display high levels of CDK11 expression. CDK11 expression knocked down by either lentiviral shRNA or siRNA inhibit cell growth and induce apoptosis in osteosarcoma cells. Immunohistochemical analysis showed that patients with osteosarcoma with high CDK11 tumor expression levels were associated with significantly shorter survival than patients with osteosarcoma with low level of tumor CDK11 expression. Systemic in vivo administration of in vivo ready siRNA of CDK11 reduced the tumor growth in an osteosarcoma subcutaneous xenograft model.Conclusions: We show that CDK11 signaling is essential in osteosarcoma cell growth and survival, further elucidating the regulatory mechanisms controlling the expression of CDK11 and ultimately develop a CDK11 inhibitor that may provide therapeutic benefit against osteosarcoma. Clin Cancer Res; 18(17); 4580-8. (C) 2012 AACR.