The Antitumoral/Antimetastatic Action of the Flavonoid Brachydin A in Metastatic Prostate Tumor Spheroids In Vitro Is Mediated by (Parthanatos) PARP-Related Cell Death
Diego Luis Ribeiro,
Katiuska Tuttis,
Larissa Cristina Bastos de Oliveira,
Juliana Mara Serpeloni,
Izabela Natalia Faria Gomes,
André van Helvoort Lengert,
Cláudia Quintino da Rocha,
Rui Manuel Reis,
Ilce Mara de Syllos Cólus,
Lusânia Maria Greggi Antunes
Affiliations
Diego Luis Ribeiro
Departament of Genetics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil
Katiuska Tuttis
Departament of Genetics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil
Larissa Cristina Bastos de Oliveira
Department of General Biology, Center for Biological Sciences, State University of Londrina, Londrina 86057-970, Brazil
Juliana Mara Serpeloni
Department of General Biology, Center for Biological Sciences, State University of Londrina, Londrina 86057-970, Brazil
Izabela Natalia Faria Gomes
Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil
André van Helvoort Lengert
Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil
Cláudia Quintino da Rocha
Department of Chemistry, Center for Exact Sciences and Technology, Federal University of Maranhão, São Luís 65080-805, Brazil
Rui Manuel Reis
Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos 14784-400, Brazil
Ilce Mara de Syllos Cólus
Department of General Biology, Center for Biological Sciences, State University of Londrina, Londrina 86057-970, Brazil
Lusânia Maria Greggi Antunes
Department of Clinical Analysis, Toxicology, and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto 14040-903, Brazil
Metastatic prostate cancer (mPCa) is resistant to several chemotherapeutic agents. Brachydin A (BrA), a glycosylated flavonoid extracted from Fridericia platyphylla, displays a remarkable antitumoral effect against in vitro mPCa cells cultured as bidimensional (2D) monolayers. Considering that three-dimensional (3D) cell cultures provide a more accurate response to chemotherapeutic agents, this study investigated the antiproliferative/antimetastatic effects of BrA and the molecular mechanisms underlying its action in mPCa spheroids (DU145) in vitro. BrA at 60–100 μM was cytotoxic, altered spheroid morphology/volume, and suppressed cell migration and tumor invasiveness. High-content analysis revealed that BrA (60–100 µM) reduced mitochondrial membrane potential and increased apoptosis and necrosis markers, indicating that it triggered cell death mechanisms. Molecular analysis showed that (i) 24-h treatment with BrA (80–100 µM) increased the protein levels of DNA disruption markers (cleaved-PARP and p-γ-H2AX) as well as decreased the protein levels of anti/pro-apoptotic (BCL-2, BAD, and RIP3K) and cell survival markers (p-AKT1 and p-44/42 MAPK); (ii) 72-h treatment with BrA increased the protein levels of effector caspases (CASP3, CASP7, and CASP8) and inflammation markers (NF-kB and TNF-α). Altogether, our results suggest that PARP-mediated cell death (parthanatos) is a potential mechanism of action. In conclusion, BrA confirms its potential as a candidate drug for preclinical studies against mPCa.
