Laminin peptides inducing invadopodia in a cell line derived from squamous cell carcinoma: dynamic study by 4D microscopy

• Grant number: 09/17336-6
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: March 01, 2010
• End date: February 28, 2014
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Ruy Gastaldoni Jaeger
• Grantee: Adriane Sousa de Siqueira
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Metastatic cells rely on invadopodia to degrade and invade extracellular matrix (ECM). Invadopodia are membrane protrusions with enzymes required for ECM degradation. These protrusions contain cortactin and membrane type 1 matrix metalloproteinase (MT1-MMP) superimposed to areas of digested matrix. This project will address invadopodia dynamics in a cell line (OSCC) derived from human oral squamous cell carcinoma. OSCC cells will be treated by laminin-derived peptides. The rationale to use these peptides is their importance in carcinoma biology, as shown previously by our group (Freitas et al., 2004; Freitas et al., 2007; Morais Freitas et al., 2007; Gama-de-Souza, 2008; Gama-de-Souza et al., 2008; Siqueira et al., 2009). OSCC cells will be treated with laminin peptides and subjected to fluorescent-substrate degradation assay. In this assay, cells are grown on fluorescent substrate. Invadopodia activity is characterized by digestion spots in fluorescent substrate, appearing as black areas in green background. Morphometry of digestion spots will inform whether laminin peptides would have induced invadopodia formation. Invadopodia dynamics will be evaluated. OSCC cells will be transfected with vectors expressing invadopodia proteins, such as cortactin-GFP and mCherry-actin, followed by treatment with laminin peptides and growth on fluorescent substrate. Invadopodia dynamics will be analyzed by 4D microscopy of living cells, collecting Z sections at different time points. 4D data will be three-dimensionally reconstructed and volumetrically rendered. Receptors and signaling pathways involved in peptide-induced invadopodia formation will be also investigated. OSCC cells will have either integrins or syndecan-1 silenced by siRNA, followed by treatment with laminin peptides and growth on fluorescent substrate. Furthermore, cells will be treated with signaling pathway inhibitors, followed by treatment with laminin peptides and growth on fluorescent substrate. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CARREIRA FALCAO, ALINE SEMBLANO; DA SILVA KATAOKA, MARIA SUELI; BAILAO RIBEIRO, NELSON ANTONIO; PICANCO DINIZ, JR., JOSE ANTONIO; ALVES, JR., SERGIO MELO; RIBEIRO RIBEIRO, ANDRE L.; DE SIQUEIRA, ADRIANE SOUSA; DA SILVA, ARTUR LUIZ; JUCA RAMOS, ROMMEL THIAGO; FREITAS, VANESSA M.; et al. A Novel Cell Line Derived from Pleomorphic Adenoma Expresses MMP2, MMP9, TIMP1, TIMP2, and Shows Numeric Chromosomal Anomalies. PLoS One, v. 9, n. 8, AUG 19 2014. (09/17336-6)

NASCIMENTO, CAMILA F.; DE SIQUEIRA, ADRIANE S.; PINHEIRO, JOAO J. V.; FREITAS, VANESSA M.; JAEGER, RUY G.. Laminin-111 derived peptides AG73 and C16 regulate invadopodia activity of a human adenoid cystic carcinoma cell line. Experimental Cell Research, v. 317, n. 18, p. 2562-2572, NOV 1 2011. (09/17336-6)

PINHEIRO DA ROSA, MARINA ROLO; CARREIRA FALCAO, ALINE SEMBLANO; FUZII, HELLEN THAIS; DA SILVA KATAOKA, MARIA SUELI; RIBEIRO, ANDRE L. R.; BOCCARDO, ENRIQUE; DE SIQUEIRA, ADRIANE SOUSA; JAEGER, RUY G.; VIANA PINHEIRO, JOAO DE JESUS; ALVES JUNIOR, SERGIO DE MELO. EGFR signaling downstream of EGF regulates migration, invasion, and MMP secretion of immortalized cells derived from human ameloblastoma. TUMOR BIOLOGY, v. 35, n. 11, p. 11107-11120, NOV 2014. (09/17336-6)

CAIRES-DOS-SANTOS, LIVIA; DA SILVA, SUELY V.; SMUCZEK, BASILIO; DE SIQUEIRA, ADRIANE S.; CRUZ, KAREN S. P.; BARBUTO, JOSE ALEXANDRE M.; AUGUSTO, TAIZE M.; FREITAS, VANESSA M.; CARVALHO, HERNANDES F.; JAEGER, RUY G.. Laminin-derived peptide C16 regulates Tks expression and reactive oxygen species generation in human prostate cancer cells. Journal of Cellular Physiology, v. 235, n. 1, JUN 2019. (09/16150-6, 18/03528-0, 09/17336-6, 10/07699-1, 15/03393-9)

SIQUEIRA, ADRIANE S.; PINTO, MONIQUE P.; CRUZ, MARIO C.; SMUCZEK, BASILIO; CRUZ, KAREN S. P.; BARBUTO, JOSE ALEXANDRE M.; HOSHINO, DAISUKE; WEAVER, ALISSA M.; FREITAS, VANESSA M.; JAEGER, RUY G.. Laminin-111 peptide C16 regulates invadopodia activity of malignant cells through beta 1 integrin, Src and ERK 1/2. ONCOTARGET, v. 7, n. 30, p. 47904-47917, JUL 26 2016. (09/17336-6)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SIQUEIRA, Adriane Sousa de. Laminin-derived peptide C16 regulates invasion and invadopodia activity/dynamics in squamous cell carcinoma and fibrosarcoma cell lines.. 2014. Doctoral Thesis - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.