Department of Radiation Oncology, The First Affiliated Hospital of China Medical University, China.
Department of Pathology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, China.
Life Sci. 2021 Apr 15;271:119148. doi: 10.1016/j.lfs.2021.119148. Epub 2021 Feb 2.
Abnormally amplified expression of AURKA (aurora kinase A) is closely related to chemo-resistance in human colorectal cancer, lung cancer and leukemia. However, the biological role of AURKA in response to radio-sensitivity in human colorectal cancer is still unknown. Therefore, we evaluated the radio-sensitize ability of perturbation AURKA in human colorectal cancer.
The knockdown effect of shAURKA was determined by western blot and qRT-PCR, respectively. Cell growth was determined by CCK-8 and clonogenic assay. Cell migration and metastasis was measured by wound healing assay and transwell invasive assay, respectively. Cell cycle and apoptosis was analyzed by flow cytometry. The alteration of down-stream targets was determined by western blot analysis.
We observed that high-level of AURKA expression is associated with poor prognosis in CRC patients receiving radiotherapy. Knockdown of AURKA significantly sensitizes the efficacy of radiation on the proliferation of HCT116 and HT-29 cells. The combination of AURKA inhibition and radiation could effectively suppress the ability of cell migration and metastasis, but also synergistically induce cellular apoptosis and arrest cell cycle at G2/M phase. Further studies demonstrated that knockdown AURKA markedly enhanced the efficacy of radiation through elevated PARP cleavage and induced AURKA-mediated pro-apoptosis factor BIM. Meanwhile, knockdown of AURKA in combination with radiation synergistically suppressed the regulator in blockage of G2/M phase, CDK2.
Taken together, our results provide the evidence that targeted inhibition of AURKA could be a promising strategy for enhancing the efficacy of radiation for the treatment of human colorectal cancer.
异常扩增的 AURKA（极光激酶 A）表达与人结直肠癌、肺癌和白血病的化疗耐药密切相关。然而，AURKA 在人结直肠癌中对放射敏感性的生物学作用尚不清楚。因此，我们评估了干扰 AURKA 对人结直肠癌放射敏感性的能力。
通过 Western blot 和 qRT-PCR 分别确定 shAURKA 的敲低效果。通过 CCK-8 和集落形成实验测定细胞生长。通过划痕愈合实验和 Transwell 侵袭实验分别测量细胞迁移和转移。通过流式细胞术分析细胞周期和凋亡。通过 Western blot 分析测定下游靶标的变化。
我们观察到 AURKA 高表达与人结直肠癌患者接受放疗后预后不良有关。敲低 AURKA 可显著增强 HCT116 和 HT-29 细胞对辐射的增殖抑制作用。AURKA 抑制联合辐射可有效抑制细胞迁移和转移能力，还可协同诱导细胞凋亡并将细胞周期阻滞在 G2/M 期。进一步的研究表明，敲低 AURKA 通过提高 PARP 切割和诱导 AURKA 介导的促凋亡因子 BIM 显著增强了辐射的疗效。同时，敲低 AURKA 联合辐射协同抑制了 G2/M 期阻滞的调节剂 CDK2。
综上所述，我们的研究结果为靶向抑制 AURKA 可能为增强人结直肠癌放射治疗的疗效提供了依据。
