Shmakova Anna A, Klimovich Polina S, Rysenkova Karina D, Popov Vladimir S, Gorbunova Anna S, Karpukhina Anna A, Karagyaur Maxim N, Rubina Kseniya A, Tkachuk Vsevolod A, Semina Ekaterina V
National Cardiology Research Center of the Ministry of Health of the Russian Federation, Institute of Experimental Cardiology, 121552 Moscow, Russia.
Faculty of Medicine, Lomonosov Moscow State University, 119192 Moscow, Russia.
Cancers (Basel). 2022 Feb 16;14(4):994. doi: 10.3390/cancers14040994.
uPAR is a membrane receptor that binds extracellular protease urokinase, contributes to matrix remodeling and plays a crucial role in cellular adhesion, proliferation, survival, and migration. uPAR overexpression in tumor cells promotes mitogenesis, opening a prospective avenue for targeted therapy. However, uPAR targeting in cancer has potential risks. We have recently shown that uPAR downregulation in neuroblastoma promotes epithelial-mesenchymal transition (EMT), potentially associated with metastasis and chemoresistance. We used data mining to evaluate the role of uPAR expression in primary and relapsed human neuroblastomas. To model the decreased uPAR expression, we targeted uPAR using CRISPR/Cas9 and shRNA in neuroblastoma Neuro2a cells and evaluated their chemosensitivity in vitro as well as tumor growth and metastasis in vivo. We demonstrate that the initially high expression predicts poor survival in human neuroblastoma. However, relapsed neuroblastomas have a significantly decreased expression. uPAR targeting in neuroblastoma Neuro2a cells leads to p38 activation and an increased p21 expression (suggesting a dormant phenotype). The dormancy in neuroblastoma cells can be triggered by the disruption of uPAR-integrin interaction. uPAR-deficient cells are less sensitive to cisplatin and doxorubicin treatment and exhibit lower p53 activation. Finally, low uPAR-expressing Neuro2a cells formed smaller primary tumors, but more frequent metastasis in mice. To the best of our knowledge, this is the first study revealing the pathological role of dormant uPAR-deficient cancer cells having a chemoresistant and motile phenotype.
尿激酶型纤溶酶原激活物受体（uPAR）是一种膜受体，可结合细胞外蛋白酶尿激酶，有助于基质重塑，并在细胞黏附、增殖、存活和迁移中发挥关键作用。肿瘤细胞中uPAR的过表达促进有丝分裂，为靶向治疗开辟了一条前景广阔的途径。然而，针对癌症的uPAR靶向治疗存在潜在风险。我们最近发现，神经母细胞瘤中uPAR的下调促进上皮-间质转化（EMT），这可能与转移和化疗耐药性有关。我们使用数据挖掘来评估uPAR表达在原发性和复发性人类神经母细胞瘤中的作用。为了模拟uPAR表达降低的情况，我们在神经母细胞瘤Neuro2a细胞中使用CRISPR/Cas9和短发夹RNA（shRNA）靶向uPAR，并评估它们在体外的化学敏感性以及在体内的肿瘤生长和转移情况。我们证明，最初的高表达预示着人类神经母细胞瘤患者的生存率较低。然而，复发性神经母细胞瘤的uPAR表达显著降低。在神经母细胞瘤Neuro2a细胞中靶向uPAR会导致p38激活和p21表达增加（提示一种休眠表型）。神经母细胞瘤细胞中的休眠可由uPAR-整合素相互作用的破坏触发。缺乏uPAR的细胞对顺铂和阿霉素治疗的敏感性较低，并且p53激活水平较低。最后，低uPAR表达的Neuro2a细胞在小鼠体内形成的原发性肿瘤较小，但转移更频繁。据我们所知，这是第一项揭示具有化疗耐药和迁移表型的休眠性uPAR缺陷癌细胞病理作用的研究。
