Department of Genetics, School of Medicine of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto, SP 14048-900, Brazil.
J Cancer Res Clin Oncol. 2012 Mar;138(3):405-14. doi: 10.1007/s00432-011-1111-0. Epub 2011 Dec 9.
Glioblastoma remains one of the most devastating human malignancies, and despite therapeutic advances, there are no drugs that significantly improve the patient survival. Altered expression of the Aurora kinases was found in different malignancies, and their inhibition has been studied in cancer therapy. In this study, we analyzed the expression of Aurora A and Aurora B in glioblastoma samples and also analyzed whether the effects of Aurora kinase inhibition were associated with temozolomide or not on cell lines and primary cultures of glioblastoma.
RT-PCR assays were used to determine the mRNA expression in glioblastoma tumor samples and in the cell lines. Cell proliferation was measured by XTT assay, and apoptosis was determined by flow cytometry. Drug combination analyses were made based in Chou-Talalay method. Gamma radiation for clonogenic survival used the doses of 2, 4 and 6 Gy. Changes in Aurora B level were assessed by Western blot analysis.
Aurora A and B were expressed in glioblastoma samples as well as in the glioblastoma cell lines (n = 6). Moreover, ZM447439, a selective Aurora kinase inhibitor, decreased the proliferation separately and synergistically with temozolomide in primary cultures and cell lines of glioblastoma. ZM also enhanced the effects of radiation on the two cell lines studied (U343 and U251), mainly when associated with TMZ in U343 cells. Treatment with ZM induced apoptotic cell death and diminished Aurora B protein level.
These data suggest that Aurora kinase inhibition may be a target for glioblastoma treatment and could be used as adjuvant to chemo- and radiotherapy.
胶质母细胞瘤仍然是最具破坏性的人类恶性肿瘤之一，尽管治疗取得了进展，但仍没有药物能显著提高患者的生存率。在不同的恶性肿瘤中发现了极光激酶的表达改变，并且已经在癌症治疗中研究了它们的抑制作用。在这项研究中，我们分析了胶质母细胞瘤样本中极光激酶 A 和极光激酶 B 的表达情况，还分析了极光激酶抑制作用的效果是否与替莫唑胺有关，无论是在细胞系还是胶质母细胞瘤的原代培养物中。
使用 RT-PCR 测定胶质母细胞瘤肿瘤样本和细胞系中的 mRNA 表达。通过 XTT 测定法测定细胞增殖，通过流式细胞术测定细胞凋亡。根据 Chou-Talalay 方法进行药物组合分析。用于集落形成存活的伽马辐射使用 2、4 和 6 Gy 的剂量。通过 Western blot 分析评估 Aurora B 水平的变化。
Aurora A 和 B 在胶质母细胞瘤样本以及胶质母细胞瘤细胞系中均有表达（n = 6）。此外，选择性 Aurora 激酶抑制剂 ZM447439 分别和与替莫唑胺联合使用，在胶质母细胞瘤的原代培养物和细胞系中均能协同抑制增殖。ZM 还增强了两种研究细胞系（U343 和 U251）对辐射的作用，主要是在 U343 细胞中与 TMZ 联合使用时。ZM 处理诱导细胞凋亡和降低 Aurora B 蛋白水平。
这些数据表明，极光激酶抑制可能是胶质母细胞瘤治疗的一个靶点，并可作为化疗和放疗的辅助手段。
