Alburquerque-González Begoña, Bernabé-García Ángel, Bernabé-García Manuel, Ruiz-Sanz Javier, López-Calderón Fernando Feliciano, Gonnelli Leonardo, Banci Lucia, Peña-García Jorge, Luque Irene, Nicolás Francisco José, Cayuela-Fuentes María Luisa, Luchinat Enrico, Pérez-Sánchez Horacio, Montoro-García Silvia, Conesa-Zamora Pablo
Department of Pathology and Histology, Campus de los Jerónimos, UCAM Universidad Católica San Antonio de Murcia, s/n, 30107 Murcia, Spain.
Laboratorio de Regeneración, Oncología Molecular y TGF-ß, Biomedical Research Institute of Murcia (IMIB-Arrixaca), Carretera Madrid-Cartagena, El Palmar, 30120 Murcia, Spain.
Cancers (Basel). 2021 Feb 18;13(4):861. doi: 10.3390/cancers13040861.
Fascin1 is the key actin-bundling protein involved in cancer invasion and metastasis whose expression is associated with bad prognosis in tumor from different origins.
In the present study, virtual screening (VS) was performed for the search of Fascin1 inhibitors and RAL, an FDA-approved inhibitor of human immunodeficiency virus-1 (HIV-1) integrase, was identified as a potential Fascin1 inhibitor. Biophysical techniques including nuclear magnetic resonance (NMR) and differential scanning fluorimetry (DSF) were carried out in order to confirm RAL as a Fascin1 blocker. The effect of RAL on actin-bundling activity Fascin1 was assessed by transmission electron microscopy (TEM), immunofluorescence, migration, and invasion assays on two human colorectal adenocarcinoma cell lines: HCT-116 and DLD-1. In addition, the anti-metastatic potential of RAL was in vivo evaluated by using the zebrafish animal model.
NMR and DSF confirmed in silico predictions and TEM demonstrated the RAL-induced disorganization of the actin structure compared to control conditions. The protrusion of lamellipodia in cancer cell line overexpressing Fascin1 (HCT-116) was abolished in the presence of this drug. By following the addition of RAL, migration of HCT-116 and DLD-1 cell lines was significantly inhibited. Finally, using endogenous and exogenous models of Fascin1 expression, the invasive capacity of colorectal tumor cells was notably impaired in the presence of RAL in vivo assays; without undesirable cytotoxic effects.
The current data show the in vitro and in vivo efficacy of the antiretroviral drug RAL in inhibiting human colorectal cancer cells invasion and metastasis in a Fascin1-dependent manner.
Fascin1是参与癌症侵袭和转移的关键肌动蛋白捆绑蛋白，其表达与不同起源肿瘤的不良预后相关。
在本研究中，进行了虚拟筛选（VS）以寻找Fascin1抑制剂，并且鉴定出FDA批准的人免疫缺陷病毒1（HIV-1）整合酶抑制剂RAL作为潜在的Fascin1抑制剂。进行了包括核磁共振（NMR）和差示扫描荧光法（DSF）在内的生物物理技术，以确认RAL是Fascin1阻断剂。通过透射电子显微镜（TEM）、免疫荧光、迁移和侵袭试验，在两种人结肠直肠腺癌细胞系HCT-116和DLD-1上评估RAL对Fascin1肌动蛋白捆绑活性的影响。此外，通过使用斑马鱼动物模型在体内评估RAL的抗转移潜力。
NMR和DSF证实了计算机模拟预测，并且TEM表明与对照条件相比，RAL诱导肌动蛋白结构紊乱。在该药物存在下，过表达Fascin1的癌细胞系（HCT-116）中片状伪足的突出被消除。随着RAL的添加，HCT-116和DLD-1细胞系的迁移受到显著抑制。最后，使用Fascin1表达的内源性和外源性模型，在体内试验中，在RAL存在下结肠直肠肿瘤细胞的侵袭能力明显受损；且无不良细胞毒性作用。
目前的数据表明抗逆转录病毒药物RAL在体外和体内以Fascin1依赖性方式抑制人结肠直肠癌细胞侵袭和转移的功效。
