Ribeiro Diego Luis, Tuttis Katiuska, Oliveira Larissa Cristina Bastos de, Serpeloni Juliana Mara, Gomes Izabela Natalia Faria, Lengert André van Helvoort, Rocha Cláudia Quintino da, Reis Rui Manuel, Cólus Ilce Mara de Syllos, Antunes Lusânia Maria Greggi
Departament of Genetics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil.
Department of General Biology, Center for Biological Sciences, State University of Londrina, Londrina 86057-970, Brazil.
Pharmaceutics. 2022 Apr 29;14(5):963. doi: 10.3390/pharmaceutics14050963.
Metastatic prostate cancer (mPCa) is resistant to several chemotherapeutic agents. Brachydin A (BrA), a glycosylated flavonoid extracted from , 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.
转移性前列腺癌（mPCa）对多种化疗药物具有抗性。从[具体来源未给出]中提取的糖基化黄酮类化合物短叶苏木酚A（BrA），对二维（2D）单层培养的体外mPCa细胞显示出显著的抗肿瘤作用。鉴于三维（3D）细胞培养对化疗药物的反应更准确，本研究调查了BrA在体外对mPCa球体（DU145）的抗增殖/抗转移作用及其作用的分子机制。60 - 100 μM的BrA具有细胞毒性，改变了球体形态/体积，并抑制了细胞迁移和肿瘤侵袭性。高内涵分析表明，BrA（60 - 100 µM）降低了线粒体膜电位，增加了凋亡和坏死标志物，表明其触发了细胞死亡机制。分子分析显示：（i）用BrA（80 - 100 µM）处理24小时可增加DNA破坏标志物（裂解的PARP和p-γ-H2AX）的蛋白质水平，同时降低抗凋亡/促凋亡（BCL-2、BAD和RIP3K）及细胞存活标志物（p-AKT1和p-44/42 MAPK）的蛋白质水平；（ii）用BrA处理72小时可增加效应半胱天冬酶（CASP3、CASP7和CASP8）和炎症标志物（NF-κB和TNF-α）的蛋白质水平。总之，我们的结果表明PARP介导的细胞死亡（副凋亡）是一种潜在的作用机制。综上所述，BrA证实了其作为抗mPCa临床前研究候选药物的潜力。
