Department of Medicine, Tulane University School of Medicine, New Orleans, LA, USA.
Department of Pathology, Tulane University School of Medicine, New Orleans, LA, USA.
Breast Cancer Res Treat. 2021 Aug;189(1):49-61. doi: 10.1007/s10549-021-06295-4. Epub 2021 Jul 1.
Breast cancer remains a prominent global disease affecting women worldwide despite the emergence of novel therapeutic regimens. Metastasis is responsible for most cancer-related deaths, and acquisition of a mesenchymal and migratory cancer cell phenotypes contributes to this devastating disease. The utilization of kinase targets in drug discovery have revolutionized the field of cancer research but despite impressive advancements in kinase-targeting drugs, a large portion of the human kinome remains understudied in cancer. NEK5, a member of the Never-in-mitosis kinase family, is an example of such an understudied kinase. Here, we characterized the function of NEK5 in breast cancer.
Stably overexpressing NEK5 cell lines (MCF7) and shRNA knockdown cell lines (MDA-MB-231, TU-BcX-4IC) were utilized. Cell morphology changes were evaluated using immunofluorescence and quantification of cytoskeletal components. Cell proliferation was assessed by Ki-67 staining and transwell migration assays tested cell migration capabilities. In vivo experiments with murine models were necessary to demonstrate NEK5 function in breast cancer tumor growth and metastasis.
NEK5 activation altered breast cancer cell morphology and promoted cell migration independent of effects on cell proliferation. NEK5 overexpression or knockdown does not alter tumor growth kinetics but promotes or suppresses metastatic potential in a cell type-specific manner, respectively.
While NEK5 activity modulated cytoskeletal changes and cell motility, NEK5 activity affected cell seeding capabilities but not metastatic colonization or proliferation in vivo. Here we characterized NEK5 function in breast cancer systems and we implicate NEK5 in regulating specific steps of metastatic progression.
尽管出现了新的治疗方案，但乳腺癌仍然是一种影响全球女性的突出的全球性疾病。转移是导致大多数癌症相关死亡的原因，而获得间充质和迁移癌细胞表型有助于这种毁灭性疾病的发生。在药物发现中利用激酶靶标已经彻底改变了癌症研究领域，但尽管在激酶靶向药物方面取得了令人印象深刻的进展，但人类激酶组的很大一部分在癌症中仍未得到充分研究。NEK5 是丝氨酸/苏氨酸激酶家族中的一个成员，是一个研究不足的激酶的例子。在这里，我们描述了 NEK5 在乳腺癌中的功能。
使用稳定过表达 NEK5 的细胞系（MCF7）和 shRNA 敲低细胞系（MDA-MB-231、TU-BcX-4IC）。通过免疫荧光和细胞骨架成分的定量评估细胞形态变化。通过 Ki-67 染色评估细胞增殖，通过 Transwell 迁移实验测试细胞迁移能力。需要进行小鼠模型的体内实验来证明 NEK5 在乳腺癌肿瘤生长和转移中的功能。
NEK5 的激活改变了乳腺癌细胞的形态并促进了细胞迁移，而与细胞增殖的影响无关。NEK5 的过表达或敲低不会改变肿瘤生长动力学，但以细胞类型特异性的方式分别促进或抑制转移潜能。
虽然 NEK5 活性调节细胞骨架变化和细胞迁移，但 NEK5 活性影响细胞播种能力，但不影响体内的转移定植或增殖。在这里，我们描述了 NEK5 在乳腺癌系统中的功能，并暗示 NEK5 参与调节转移进展的特定步骤。
