ASPM介导的AURKA表达和活性调节在胶质瘤Epothilone B化疗抵抗中的作用及机制研究

Glioma is the most common primary malignant tumor of the central nervous system. However, the molecular mechanism of the progression and chemotherapy resistance of glioma remains unclear. In our previous study, we found that human abnormal spindle microcephaly related protein (ASPM) was involved in the malignant proliferation and migration of glioma cells, and the expression of ASPM in glioma cells was positively correlated with Epothilone B sensitivity. In addition, recent studies found that ASPM can bind with AURKA, and our previous studies found that AURKA expression in glioma tissues is significantly positively correlated with ASPM expression. In U87-MG cells, inhibition of ASPM could significantly down-regulate AURKA protein expression and increase the Epothilone B sensitivity. Therefore, we hypothesize that ASPM may affect the sensitivity of Epothilone B by regulating the expression and activity of AURKA in glioma. Here, we plan to identify the correlation between ASPM and AURKA expression and the prognosis of glioma patients and Epothilone B sensitivity of glioma cells. Coimmunoprecipitation assay and GST-pulldown assay will be used to identify the interaction between ASPM and AURKA protein in glioma cells. Finally, cell and animal experiments will be applied to clarify the role of ASPM-mediated AURKA expression and activity regulation in Epothilone B resistance of glioma. This study will contribute to establish new potentially targets for glioma therapy and provide new insight for individualized treatment of glioma with Epothilone B resistance.

脑胶质瘤是最常见的中枢神经系统原发性恶性肿瘤，因其侵袭性强和放化疗抵抗的特点，患者预后差。本项目前期实验发现人类异常纺锤体样小头畸形相关蛋白（ASPM）可促进胶质瘤细胞增殖和迁移；文献报道ASPM可与激酶AURKA结合，而我们发现胶质瘤组织中AURKA与ASPM表达呈正相关，干扰胶质瘤细胞中ASPM表达可下调AURKA蛋白表达，增加Epothilone B（EpoB）敏感性。因此本项目拟通过临床研究，查明ASPM和AURKA表达与胶质瘤患者预后及胶质瘤细胞EpoB敏感性的相关性；并应用蛋白免疫共沉淀和GST-pulldown实验，查明胶质瘤细胞中ASPM与AURKA蛋白相互作用；最后通过细胞和动物实验，阐明ASPM介导的AURKA表达和活性调节在胶质瘤EpoB化疗抵抗中的作用。本研究有助于为胶质瘤的治疗提供新的靶点，并为基于ASPM表达水平胶质瘤患者EpoB的个体化治疗奠定重要的理论依据。

全基因组表达谱分析为探索肿瘤发生发展的机制提供了新的方法，可以有效识别参与肿瘤发生发展的致病基因。本项目通过生物信息学分析和临床样本验证发现ASPM在胶质瘤组织中异常高表达并提示患者较差的预后；干扰ASPM可以抑制胶质瘤细胞的增殖、迁移，诱导细胞周期阻滞，减缓胶质瘤裸鼠皮下肿瘤的生长；并首次揭示ASPM介导AURKA表达是调控胶质瘤发生发展的关键环节。Epothilone B 在胶质瘤治疗中具有良好的前景，在广泛的临床应用前揭示其疗效个体差异或抵抗机制对指导该药的临床试验或应用具有重要意义。本项目首次发现干预 ASPM 表达可以增加胶质瘤细胞对 Epothilone B 化疗敏感性，并提出ASPM可能通过影响AURKA介导 Epothilone B 化疗抵抗的分子机制。本研究可以为胶质瘤的个体化治疗提供科学依据，同时为Epothilone B 应用于治疗胶质瘤以及逆转 Epothilone B 化疗抵抗提供重要的药理学依据和思路。