Begoña Alburquerque-González1, Ángel Bernabé-García2, Manuel Bernabé-García3, Javier Ruiz-Sanz4, Fernando Feliciano López-Calderón1, Leonardo Gonnelli5,6, Lucia Banci5,6, Jorge Peña-García7, Irene Luque4, Francisco José Nicolás2, María Luisa Cayuela-Fuentes3, Enrico Luchinat5,8, Horacio Pérez-Sánchez7, Silvia Montoro-García9, Pablo Conesa-Zamora10. 1. Department of Pathology and Histology, Campus de los Jerónimos, UCAM Universidad Católica San Antonio de Murcia, s/n, 30107 Murcia, Spain. 2. Laboratorio de Regeneración, Oncología Molecular y TGF-ß, Biomedical Research Institute of Murcia (IMIB-Arrixaca), Carretera Madrid-Cartagena, El Palmar, 30120 Murcia, Spain. 3. Telomerase, Cancer and Aging Group, Biomedical Research Institute of Murcia (IMIB-Arrixaca), 30120 Murcia, Spain. 4. Department of Physical Chemistry, Institute of Biotechnology and Excellence Research Unit of "Chemistry Applied to Biomedicine and the Environment, Spain Campus Fuentenueva s/n, University of Granada, 18071 Granada, Spain. 5. CERM-Magnetic Resonance Center, Università degli Studi di Firenze, Via Luigi Sacconi 6, 50019 Sesto Fiorentino, Italy. 6. Dipartimento di Chimica, Università degli Studi di Firenze, Via della Lastruccia 3, 50019 Sesto Fiorentino, Italy. 7. Structural Bioinformatics and High Performance Computing (BIO-HPC) Research Group, Campus de los Jerónimos, s/n, UCAM Universidad Católica San Antonio de Murcia, Guadalupe, 30107 Murcia, Spain. 8. Consorzio per lo Sviluppo dei Sistemi a Grande Interfase-CSGI, Via della Lastruccia 3, 50019 Sesto Fiorentino, Italy. 9. Cell Culture Lab, Facultad de Ciencias de la Salud, Campus de los Jerónimos, s/n, UCAM Universidad Católica San Antonio de Murcia, Guadalupe, 30107 Murcia, Spain. 10. Laboratory Medicine Department, Group of Molecular Pathology and Pharmacogenetics, Biomedical Research Institute from Murcia (IMIB), Hospital Universitario Santa Lucía, c/Mezquita sn, 30202 Cartagena, Spain.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
BACKGROUND: 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. METHODS: In the present study, virtual screening (VS) was performed for the search of Fascin1 inhibitors and RAL, an FDA-approved inhibitor of humanimmunodeficiency 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 humancolorectal 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. RESULTS: 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. CONCLUSION: The current data show the in vitro and in vivo efficacy of the antiretroviral drug RAL in inhibiting humancolorectal cancer cells invasion and metastasis in a Fascin1-dependent manner.
Authors: Shengchen Lin; Yunzhan Li; Dezhen Wang; Chongbiao Huang; David Marino; Oana Bollt; Chaodong Wu; Matthew D Taylor; Wei Li; Gina M DeNicola; Jihui Hao; Pankaj K Singh; Shengyu Yang Journal: Cancer Lett Date: 2021-07-22 Impact factor: 9.756