Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
J Exp Clin Cancer Res. 2020 Sep 10;39(1):183. doi: 10.1186/s13046-020-01659-y.
NEK2, a serine/threonine kinase involved in mitosis, has been found to function in chromosome instability, tumor progression and metastasis, but its role in cervical cancer radioresistance remains unknown.
We detected the protein levels of NEK2 in cervical carcinoma tissues and paired paracarcinoma tissues by immunohistochemistry. The roles of NEK2 in oncogenesis were examined using cell growth and colony formation assays, EdU assay, apoptosis assay as well as in vivo mouse model. γ-H2AX and Rad51 foci formation, neutral comet assay and clonogenic cell survival assay were applied to determine the radiosensitivity of cervical cancer cells. RNA-seq was performed to identify the downstream effector of NEK2. The gene expression levels were measured by Real-time PCR.
We report that NEK2 protein level is overexpressed and correlated with the tumor stage and lymph node metastasis in cervical cancer tissues. Furthermore, we provided evidence that depletion of NEK2 impairs oncogenesis and enhances radiosensitivity in cervical cancer. Using RNA sequencing, we identify Wnt1 as a key downstream effector of NEK2. Knockdown of NEK2 downregulates the mRNA and protein levels of Wnt1, thereby inhibiting the activation of the Wnt/β-catenin signaling pathway. More importantly, the observed consequences induced by NEK2 depletion in cervical cancer cells can be partially rescued by Wnt1 overexpression.
Our results demonstrate that NEK2 activates the Wnt/β-catenin signaling pathway via Wnt1 to drive oncogenesis and radioresistance in cervical cancer, indicating that NEK2 may be a promising target for the radiosensitization of cervical cancer.
参与有丝分裂的丝氨酸/苏氨酸激酶 NEK2 已被发现在染色体不稳定性、肿瘤进展和转移中发挥作用，但它在宫颈癌放射抵抗中的作用尚不清楚。
我们通过免疫组织化学检测了宫颈癌组织和配对癌旁组织中 NEK2 的蛋白水平。使用细胞生长和集落形成测定、EdU 测定、凋亡测定以及体内小鼠模型来研究 NEK2 在致癌作用中的作用。γ-H2AX 和 Rad51 焦点形成、中性彗星测定和克隆形成细胞存活测定用于确定宫颈癌细胞的放射敏感性。进行 RNA-seq 以鉴定 NEK2 的下游效应子。通过 Real-time PCR 测量基因表达水平。
我们报告说，NEK2 蛋白水平在宫颈癌组织中过度表达，并与肿瘤分期和淋巴结转移相关。此外，我们提供的证据表明，NEK2 的缺失会损害宫颈癌的发生，并增强其放射敏感性。通过 RNA 测序，我们确定 Wnt1 是 NEK2 的关键下游效应子。NEK2 的敲低下调了 Wnt1 的 mRNA 和蛋白水平，从而抑制了 Wnt/β-catenin 信号通路的激活。更重要的是，通过 NEK2 耗竭在宫颈癌细胞中观察到的结果可以部分通过 Wnt1 的过表达来挽救。
我们的研究结果表明，NEK2 通过 Wnt1 激活 Wnt/β-catenin 信号通路，从而驱动宫颈癌的发生和放射抵抗，表明 NEK2 可能是宫颈癌放射增敏的一个有前途的靶点。
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