Center for Sarcoma and Connective Tissue Oncology and Department of Pathology, Massachusetts General Hospital, Boston, MA 02114, USA.
Clin Cancer Res. 2012 Sep 1;18(17):4580-8. doi: 10.1158/1078-0432.CCR-12-1157. Epub 2012 Jul 12.
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.
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.
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.
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.
鉴定新的靶向治疗方法对于提高骨肉瘤患者的生存率至关重要。本研究的目的是鉴定骨肉瘤中基于激酶的潜在治疗靶点。
我们使用基于慢病毒的 shRNA 激酶文库筛选在骨肉瘤细胞存活中起作用的激酶。细胞增殖测定用于评估细胞生长和存活。siRNA 测定用于证实观察到的表型变化是由于激酶基因表达的丧失所致。CDK11（PITSLRE）被鉴定为骨肉瘤细胞存活所必需，其表达通过 Western blot 分析和免疫组织化学证实。总体患者存活率与 CDK11 表达及其预后相关。在体内骨肉瘤异种移植模型中进一步评估 CDK11 表达在维持骨肉瘤生长中的作用。
骨肉瘤细胞显示出高水平的 CDK11 表达。通过慢病毒 shRNA 或 siRNA 敲低 CDK11 表达抑制骨肉瘤细胞的生长并诱导其凋亡。免疫组织化学分析表明，CDK11 肿瘤高表达的骨肉瘤患者的生存率明显低于 CDK11 肿瘤低表达的骨肉瘤患者。体内 ready siRNA 对 CDK11 的系统体内给药减少了骨肉瘤皮下异种移植模型中的肿瘤生长。
我们表明 CDK11 信号在骨肉瘤细胞生长和存活中是必需的，进一步阐明了控制 CDK11 表达的调节机制，并最终开发出一种 CDK11 抑制剂，可能对骨肉瘤提供治疗益处。
